He wero tonu te tātaritanga rerekētanga o ngā pona pūkahukahu i tautuhia mā te tomography rorohiko (CT) i roto i ngā mahi haumanu. I konei, ka whakaahuahia e mātou te metabolome ā-ao o ngā tauira toto 480, tae atu ki ngā mana hauora, ngā pona pūkahukahu pai, me te adenocarcinoma pūkahukahu wāhanga I. He āhua motuhake ngā āhua metabolomic o ngā adenocarcinoma, engari he rite tonu ngā āhua metabolomic o ngā pona pai me ngā tāngata hauora. I roto i te rōpū kitenga (n = 306), i tautuhia he huinga o ngā metabolite 27 hei wehewehe i waenga i ngā pona pai me ngā pona kino. Ko te AUC o te tauira wehewehe i roto i ngā rōpū whakamana ā-roto (n = 104) me te whakamana ā-waho (n = 111) he 0.915 me te 0.945, ia. I whakaatuhia e te tātari ara te pikinga o ngā metabolite glycolytic e pā ana ki te hekenga o te tryptophan i roto i te toto adenocarcinoma pūkahukahu ki te whakataurite ki ngā pona pai me ngā mana hauora, ā, i kīia ko te tangohanga tryptophan ka whakatairanga i te glycolysis i roto i ngā pūtau matepukupuku pūkahukahu. E whakaatu ana tā mātou rangahau i te uara o ngā tohu koiora metabolite toto i roto i te aromatawai i te mōrearea o ngā pona pūkahukahu i kitea e te CT.
He mea nui te tautuhi wawe i te mate hei whakapai ake i ngā reiti oranga mō ngā tūroro mate pukupuku. Kua whakaatuhia e ngā hua o te US National Lung Cancer Screening Trial (NLST) me te European NELSON Study ka taea e te tirotiro me te iti-horopeta computed tomography (LDCT) te whakaiti i te mate pukupuku pūkahukahu i roto i ngā rōpū mōrearea nui1,2,3. Mai i te whakamahinga whānui o te LDCT mō te tirotiro mate pukupuku pūkahukahu, kua piki haere tonu te maha o ngā kitenga x-ray tūpono o ngā pona pūkahukahu kāore he tohu 4. Ko ngā pona pūkahukahu e tautuhia ana ko ngā opacities arotahi tae atu ki te 3 cm te whānui 5. He uaua ki a mātou te aromatawai i te tūponotanga o te mate pukupuku me te whakatau i te maha o ngā pona pūkahukahu i kitea tūpono i runga i te LDCT. Ka taea e ngā herenga o te CT te arahi ki ngā whakamātautau whai muri pinepine me ngā hua teka-pai, ka arahi ki te wawaotanga koretake me te maimoatanga nui6. Nō reira, me whakawhanake he tohu koiora pono me te whai hua hei tautuhi tika i te mate pukupuku pūkahukahu i ngā wāhanga tōmua me te wehewehe i te nuinga o ngā pona pai i te kitenga tuatahi 7.
Ko te tātaritanga whānui o te toto (te toto toto, te plasma, ngā pūtau mononuclear toto ā-taha), tae atu ki te genomics, te proteomics, te DNA methylation8,9,10 rānei, kua piki ake te hiahia ki te kitea o ngā tohu koiora mō te mate pukupuku pūkahukahu. I taua wā, ko ngā huarahi metabolomics e ine ana i ngā hua mutunga pūtau e pāngia ana e ngā mahi ā-roto me ā-waho, ā, ka whakamahia hei matapae i te tīmatanga me te putanga o te mate. Ko te wai chromatography-tandem mass spectrometry (LC-MS) he tikanga e whakamahia whānuitia ana mō ngā rangahau metabolomics nā tōna tairongo teitei me te whānuitanga hihiri nui, ka taea te kapi i ngā metabolites me ngā āhuatanga ā-tinana me ngā āhuatanga matū rerekē11,12,13. Ahakoa kua whakamahia te tātaritanga metabolomic ā-ao o te plasma/serum hei tautuhi i ngā tohu koiora e pā ana ki te tātaritanga mate pukupuku pūkahukahu14,15,16,17 me te whai huatanga o te maimoatanga,18 kei te rangahauhia tonutia ngā whakarōpūtanga metabolite serum hei wehewehe i waenga i ngā pona pūkahukahu pai me te kino. -rangahau nui.
Ko te adenocarcinoma me te squamous cell carcinoma ngā momo matua e rua o te mate pukupuku pūkahukahu kore-pūtau iti (NSCLC). He maha ngā whakamātautau tirotiro CT e tohu ana ko te adenocarcinoma te momo histological tino noa o te mate pukupuku pūkahukahu1,19,20,21. I roto i tēnei rangahau, i whakamahia e mātou te ultra-performance liquid chromatography-high-resolution mass spectrometry (UPLC-HRMS) hei whakahaere i te tātari metabolomics i runga i te katoa o te 695 tauira serum, tae atu ki ngā mana hauora, ngā pona pūkahukahu pai, me te CT-i kitea ≤3 cm. Te tirotiro mō te adenocarcinoma pūkahukahu Wāhanga I. I tautuhia e mātou he rōpū o ngā metabolites serum e wehewehe ana i te adenocarcinoma pūkahukahu mai i ngā pona pai me ngā mana hauora. I whakaatuhia e te tātari whakarei ara ko te tryptophan rerekē me te metabolism glucose he huringa noa i roto i te adenocarcinoma pūkahukahu ki te whakataurite ki ngā pona pai me ngā mana hauora. Hei whakamutunga, i whakatūhia e mātou, ā, i whakamanahia he whakarōpūtanga metabolic serum me te tino motuhake me te aro ki te wehewehe i waenga i ngā pona pūkahukahu kino me te pai i kitea e te LDCT, ka āwhina pea i te tautuhi rerekētanga wawe me te aromatawai mōrearea.
I roto i tēnei rangahau o nāianei, i kohia whakamuri ngā tauira toto toto i rite ki te ira tangata me te tau mai i te 174 mana hauora, 292 tūroro he pona pūkahukahu pai, me te 229 tūroro he adenocarcinoma pūkahukahu wāhanga I. Kei te whakaatuhia ngā āhuatanga taupori o ngā kaupapa 695 i roto i te Ripanga Tāpiri 1.
E ai ki te Pikitia 1a, e 480 ngā tauira toto, tae atu ki te 174 o ngā tauira hauora (HC), 170 ngā pona pai (BN), me te 136 o ngā tauira adenocarcinoma pūkahukahu (LA) wāhanga I, i kohia ki te Sun Yat-sen University Cancer Center. He rōpū kitenga mō te whakatauira metabolomic kore-whaiaro mā te whakamahi i te ultra-performance liquid chromatography-high-resolution mass spectrometry (UPLC-HRMS). E ai ki te Pikitia Tāpiri 1, i tautuhia ngā metabolites rerekē i waenga i te LA me te HC, LA me te BN hei whakatū i tētahi tauira whakarōpūtanga me te tūhura atu i te tātaritanga ara rerekētanga. 104 ngā tauira i kohia e te Sun Yat-sen University Cancer Center me te 111 ngā tauira i kohia e ētahi atu hōhipera e rua i whakamanahia ā-roto me ā-waho.
a Te taupori rangahau i roto i te rōpū kitenga i tirotirohia te metabolomics toto o te ao mā te whakamahi i te ultra-performance liquid chromatography-high-resolution mass spectrometry (UPLC-HRMS). b Te tātari wehewehe tapawhā iti rawa wāhanga (PLS-DA) o te metabolome katoa o te 480 tauira toto mai i te rōpū rangahau, tae atu ki ngā mana hauora (HC, n = 174), ngā pona pai (BN, n = 170), me te adenocarcinoma pūkahukahu wāhanga I (Los Angeles, n = 136). +ESI, aratau ionization electrospray pai, -ESI, aratau ionization electrospray kino. c–e Ko ngā Metabolites me ngā nui rerekē nui i roto i ngā rōpū e rua kua hoatu (whakamātautau Wilcoxon tohu rua-hiku, uara p kua whakatikatikaina te tere kitenga teka, FDR <0.05) e whakaatuhia ana i te whero (huringa takai > 1.2) me te kikorangi (huringa takai < 0.83). ) e whakaatuhia ana i runga i te whakairoiro puia. He mahere wera whakarōpū whakarōpū e whakaatu ana i ngā rerekētanga nui o te maha o ngā matū whai tohu i waenga i te LA me te BN. Ka whakaratohia ngā raraunga pūtake i roto i te ahua o ngā kōnae raraunga pūtake.
I tātarihia te metabolome toto katoa o te 174 HC, 170 BN me te 136 LA i roto i te rōpū kitenga mā te whakamahi i te tātaritanga UPLC-HRMS. I te tuatahi ka whakaaturia e mātou ko ngā tauira whakahaere kounga (QC) e huihui pūmau ana i waenganui o te tauira tātaritanga wāhanga matua (PCA) kāore i te tirotirohia, e whakaū ana i te pumau o te mahi a te rangahau o nāianei (Pikitia Tāpiri 2).
E ai ki te tātari wehewehe-tapawhā iti rawa wāhanga (PLS-DA) i te Pikitia 1 b, i kitea e mātou he mārama ngā rerekētanga i waenga i te LA me te BN, LA me te HC i roto i ngā aratau ionization electrospray pai (+ESI) me te kino (−ESI) i wehea. Heoi, kāore i kitea he rerekētanga nui i waenga i te BN me te HC i roto i ngā āhuatanga +ESI me te -ESI.
I kitea e mātou he 382 ngā āhuatanga rerekētanga i waenga i te LA me te HC, he 231 ngā āhuatanga rerekētanga i waenga i te LA me te BN, me te 95 ngā āhuatanga rerekētanga i waenga i te BN me te HC (whakamātautau Wilcoxon signed rank, FDR <0.05 me te huringa maha >1.2 <0.83 rānei) (Pikitia .1c-e). . I tāpirihia anō ngā tihi (Raraunga Tāpiri 3) ki tētahi pātengi raraunga (whare pukapuka mzCloud/HMDB/Chemspider) mā te uara m/z, te wā pupuri me te rapunga whānuitanga papatipu wehewehe (ngā taipitopito kua whakaahuahia i roto i te wāhanga Tikanga) 22 . Hei whakamutunga, 33 me te 38 ngā matū whaihua kua tāpirihia me ngā rerekētanga nui o te nui i tautuhia mō te LA ki te BN (Pikitia 1f me te Ripanga Tāpiri 2) me te LA ki te HC (Pikitia Tāpiri 3 me te Ripanga Tāpiri 2), ia. He rerekē, 3 noa iho ngā matū whaihua me ngā rerekētanga nui o te nui i tautuhia i roto i te BN me te HC (Ripanga Tāpiri 2), e rite ana ki te taupatupatu i waenga i te BN me te HC i roto i te PLS-DA. Ka kapi katoa ēnei matū rerekē i te whānuitanga o ngā matū koiora (Pikitia Tāpiri 4). Ina huihuia, e whakaatu ana ēnei hua i ngā huringa nui o te metabolome toto e whakaata ana i te huringa kino o te mate pukupuku pūkahukahu i te wā tōmua ki te whakataurite ki ngā pona pūkahukahu pai, ki ngā tāngata hauora rānei. I taua wā tonu, ko te ōrite o te metabolome toto o BN me HC e kī ana he maha ngā āhuatanga koiora o ngā pona pūkahukahu pai ki ngā tāngata hauora. I te mea he mea noa ngā whakarerekētanga ira epidermal growth factor receptor (EGFR) i roto i te momo adenocarcinoma pūkahukahu 23, i whai mātou ki te whakatau i te pānga o ngā whakarerekētanga taraiwa ki te metabolome toto. I muri mai ka tātarihia e mātou te kōtaha metabolomic whānui o te 72 ngā take me te tūnga EGFR i roto i te rōpū adenocarcinoma pūkahukahu. He mea whakamīharo, i kitea e mātou he kōtaha rite i waenga i ngā tūroro EGFR mutant (n = 41) me ngā tūroro EGFR momo-mohoao (n = 31) i roto i te tātaritanga PCA (Pikitia Tāpiri 5a). Heoi, i kitea e mātou e 7 ngā matūkai i tino rerekē te nui i roto i ngā tūroro he whakarerekētanga EGFR ki te whakaritea ki ngā tūroro he EGFR momo-mohoao (whakamātautau-t, p < 0.05 me te huringa whakapiko > 1.2, < 0.83 rānei) (Pikitia Tāpiri 5b). Ko te nuinga o ēnei matūkai (5 i roto i te 7) he acylcarnitine, he mea nui ki ngā ara waikura waikawa ngako.
E ai ki te whakaaturanga o te tukanga mahi i whakaaturia i te Pikitia 2 a, i whiwhihia ngā tohu koiora mō te whakarōpūtanga pona mā te whakamahi i ngā kaiwhakahaere whakahekenga tino iti rawa me te kōwhiringa i runga i te 33 ngā metabolite rerekē i tautuhia i roto i te LA (n = 136) me te BN (n = 170). Te huinga pai rawa atu o ngā taurangi (LASSO) – tauira whakatauranga logistic rua. I whakamahia te whakamana-whakawhiti tekau-wā hei whakamātautau i te pono o te tauira. Ka whakatikatikaina te kōwhiringa taurangi me te whakatikatika tawhā mā te whiu whakanui tūponotanga me te tawhā λ24. I whakahaerehia anō te tātari metabolomics ā-ao motuhake i roto i ngā rōpū whakamana ā-roto (n = 104) me te whakamana ā-waho (n = 111) hei whakamātautau i te mahi whakarōpūtanga o te tauira wehewehe. Nō reira, 27 ngā metabolite i roto i te huinga kitenga i tautuhia ko te tauira wehewehe pai rawa atu me te uara AUC toharite nui rawa atu (Pikitia 2b), i roto i ēnei ko te 9 he nui ake te mahi, ā, 18 he iti iho te mahi i roto i te LA ki te whakataurite ki te BN (Pikitia 2c).
Rerenga mahi mō te hanga i tētahi whakarōpūtanga pona pūkahukahu, tae atu ki te whiriwhiri i te papa pai rawa atu o ngā matū matū i roto i te huinga kitenga mā te whakamahi i tētahi tauira whakatauranga logistic rua mā te whakamana-whakawhiti tekau-whakapanga me te aromatawai i te mahi matapae i roto i ngā huinga whakamana ā-roto me ā-waho. b Ngā tatauranga whakamana-whakawhiti o te tauira whakatauranga LASSO mō te kōwhiringa tohu koiora matū. Ko ngā tau i hoatu i runga ake nei e tohu ana i te maha toharite o ngā tohu koiora i tīpakohia i tētahi λ kua hoatu. Ko te rārangi ira-whero e tohu ana i te uara AUC toharite i te lambda e rite ana. Ko ngā pae hapa hina e tohu ana i ngā uara AUC iti rawa me te mōrahi. Ko te rārangi ira-ira e tohu ana i te tauira pai rawa atu me ngā tohu koiora 27 kua tīpakohia. AUC, rohe i raro i te pihi āhuatanga whakahaere kaiwhiwhi (ROC). c Whakapikohia ngā huringa o te 27 matū kua tīpakohia i roto i te rōpū LA ki te whakataurite ki te rōpū BN i roto i te rōpū kitenga. Pou whero – whakahohenga. Ko te pou kikorangi he hekenga. d–f Ngā pihi āhuatanga whakahaere kaiwhiwhi (ROC) e whakaatu ana i te mana o te tauira wehewehe i runga i ngā huinga matū 27 i roto i ngā huinga whakamana kitenga, ā-roto, me waho. Ka whakaratohia ngā raraunga pūtake i roto i te ahua o ngā kōnae raraunga pūtake.
I hangaia he tauira matapae i runga i ngā tauwehenga whakatau taumaha o ēnei matūkai 27 (Ripanga Tāpiri 3). Ko te tātari ROC i runga i ēnei matūkai 27 i hua ake he uara rohe i raro i te pihi (AUC) o te 0.933, ko te aro o te rōpū kitenga he 0.868, ā, ko te motuhaketanga he 0.859 (Pikitia 2d). I taua wā, i roto i ngā matūkai rerekētanga 38 kua whakamāramahia i waenga i te LA me te HC, i tutuki i tētahi huinga o ngā matūkai 16 he AUC o te 0.902 me te aro o te 0.801 me te motuhaketanga o te 0.856 i te wehewehe i te LA mai i te HC (Pikitia Tāpiri 6a-c). I whakaritea hoki ngā uara AUC i runga i ngā paepae huringa papa rerekē mō ngā matūkai rerekētanga. I kitea e mātou ko te tauira whakarōpūtanga i tino pai te mahi i roto i te wehewehe i waenga i te LA me te BN (HC) i te wā i whakatakotoria ai te taumata huringa papa ki te 1.2 ki te 1.5, ki te 2.0 rānei (Pikitia Tāpiri 7a,b). I whakamanahia anō te tauira whakarōpūtanga, i runga i te 27 ngā rōpū matū matū, i roto i ngā rōpū ā-roto me ā-waho. Ko te AUC he 0.915 (aronga 0.867, motuhaketanga 0.811) mō te whakamana ā-roto me te 0.945 (aronga 0.810, motuhaketanga 0.979) mō te whakamana ā-waho (Pikitia 2e, f). Hei aromatawai i te whai huatanga o te taiwhanga, i tātarihia ngā tauira 40 mai i te rōpū ā-waho i roto i tētahi taiwhanga ā-waho e ai ki te whakaahuatanga i te wāhanga Tikanga. I tutuki i te tika o te whakarōpūtanga he AUC o te 0.925 (Pikitia Tāpiri 8). Nā te mea ko te carcinoma pūtau squamous pūkahukahu (LUSC) te tuarua o ngā momo mate pukupuku pūkahukahu kore-pūtau iti (NSCLC) i muri i te adenocarcinoma pūkahukahu (LUAD), i whakamatauria anō hoki e mātou te whai huatanga pea o ngā kōtaha matū kua whakamanahia. BN me te 16 ngā kēhi o te LUSC. Ko te AUC o te wehewehe i waenga i te LUSC me te BN he 0.776 (Pikitia Tāpiri 9), e tohu ana i te ngoikore o te kaha ki te whakataurite ki te wehewehe i waenga i te LUAD me te BN.
Kua whakaatuhia e ngā rangahau he hononga pai te rahi o ngā pona i runga i ngā whakaahua CT ki te tūponotanga o te mate pukupuku, ā, he mea nui tonu te whakatau i te maimoatanga o ngā pona25,26,27. I whakaatuhia e te tātaritanga raraunga mai i te rōpū nui o te rangahau tirotiro NELSON ko te mōrearea o te mate pukupuku i roto i ngā tāngata he pona <5 mm he rite tonu ki ērā i roto i ngā tāngata kāore he pona 28. Nō reira, ko te rahi iti rawa e hiahiatia ana te aroturuki CT auau ko te 5 mm, e ai ki te taunakitanga a te British Thoracic Society (BTS), me te 6 mm, e ai ki te taunakitanga a te Fleischner Society 29. Heoi, ko ngā pona nui ake i te 6 mm me te kore he āhuatanga pai e kitea ana, e kiia nei ko ngā pona pūkahukahu kore e taea te whakatau (IPN), he wero nui tonu i roto i te aromatawai me te whakahaere i roto i te mahi haumanu30,31. I muri mai ka tirotirohia e mātou mēnā i pā te rahi o te pona ki ngā waitohu metabolomic mā te whakamahi i ngā tauira kua kohikohia mai i ngā rōpū kitenga me te whakamana ā-roto. I te arotahi ki te 27 ngā tohu koiora kua whakamanahia, i whakaritea tuatahitia e mātou ngā kōtaha PCA o ngā metabolomes HC me BN sub-6 mm. I kitea e mātou ko te nuinga o ngā raraunga mō te HC me te BN i taupatupatu, e whakaatu ana he rite ngā taumata matūkai toto i roto i ngā rōpū e rua (Pikitia 3a). I noho pūmau ngā mahere āhuatanga puta noa i ngā awhe rahi rerekē i roto i te BN me te LA (Pikitia 3b, c), engari i kitea he wehenga i waenga i ngā pona kino me ngā pona pai i roto i te awhe 6–20 mm (Pikitia 3d). Ko te AUC o tēnei rōpū he 0.927, he motuhaketanga o te 0.868, me te aro o te 0.820 mō te matapae i te kino o ngā pona e ine ana i waenga i te 6 ki te 20 mm (Pikitia 3e, f). E whakaatu ana ā mātou hua ka taea e te kaiwhakarōpū te hopu i ngā huringa pākia i puta mai i te huringa kino wawe, ahakoa te rahi o te pona.
ad Whakataurite i ngā kōtaha PCA i waenga i ngā rōpū kua tohua i runga i te whakarōpūtanga pākia o ngā matūkai 27. CC me BN < 6 mm. b BN < 6 mm vs BN 6–20 mm. i roto i te LA 6–20 mm versus LA 20–30 mm. g BN 6–20 mm me LA 6–20 mm. GC, n = 174; BN < 6 mm, n = 153; BN 6–20 mm, n = 91; LA 6–20 mm, n = 89; LA 20–30 mm, n = 77. e Te pihi āhuatanga mahi kaiwhiwhi (ROC) e whakaatu ana i te mahi tauira wehewehe mō ngā pona 6–20 mm. f I tatauhia ngā uara tūponotanga i runga i te tauira whakatauranga logistic mō ngā pona e ine ana i te 6–20 mm. Ko te rārangi iraira hina e tohu ana i te uara tapahi tino pai (0.455). Ko ngā tau i runga ake nei e tohu ana i te ōrau o ngā take i matapaetia mō Los Angeles. Whakamahia he whakamātautau-t a te Tauira e rua-waewae. PCA, tātaritanga wāhanga matua. Te horahanga AUC i raro i te pihi. Ka whakaratohia ngā raraunga pūtake i roto i te āhua o ngā kōnae raraunga pūtake.
E whā ngā tauira (44–61 tau te pakeke) he rite te rahi o ngā pona pūkahukahu (7–9 mm) i tīpakohia anō hei whakaatu i te mahi a te tauira matapae mate pukupuku kua whakaarohia (Pikitia 4a, b). I te tirotiro tuatahi, i puta te Take 1 hei pona totoka me te whakapūmau, he āhuatanga e pā ana ki te pai, ko te Take 2 ia i puta hei pona totoka wāhanga kore e taea te whakatau me te kore he āhuatanga pai e kitea ana. E toru ngā huringa o ngā karapa CT whai muri i whakaatu i te pumau tonu o ēnei take mō te wā 4-tau, nō reira i kiia ai he pona pai (Pikitia 4a). Ki te whakatauritea ki te aromatawai haumanu o ngā karapa CT raupapa, ko te tātaritanga metabolite serum kotahi-wero me te tauira whakarōpū o nāianei i tere me te tika te tautuhi i ēnei pona pai i runga i ngā here tūponotanga (Ripanga 1). E whakaatu ana te Pikitia 4b i roto i te take 3 i tētahi pona me ngā tohu o te whakahokinga pleural, e pā ana ki te mate pukupuku32. I puta te Take 4 hei pona totoka wāhanga kore e taea te whakatau me te kore he tohu o tētahi take pai. I matapaetia ēnei kēhi katoa he mate kino e ai ki te tauira whakarōpū (Ripanga 1). I whakaaturia te aromatawai o te adenocarcinoma o te pūkahukahu mā te whakamātautau histopathological i muri i te pokanga tango pūkahukahu (Pikitia 4b). Mō te huinga whakamana o waho, i matapaetia tika e te whakarōpū metabolic ngā kēhi e rua o ngā pona pūkahukahu kore e taea te whakatau he nui ake i te 6 mm (Pikitia Tāpiri 10).
Ngā whakaahua CT o te matapihi axial o ngā pukahukahu o ngā kēhi e rua o ngā pona pai. I roto i te kēhi 1, i whakaatuhia e te matawai CT i muri i ngā tau 4 he pona totoka pumau e 7 mm te rahi me te whakapūpū i te pito matau o raro. I roto i te kēhi 2, i whakaatuhia e te matawai CT i muri i ngā tau 5 he pona pumau, he totoka wāhanga me te diameter o te 7 mm i te pito matau o runga. b Ngā whakaahua CT matapihi axial o ngā pukahukahu me ngā rangahau pathological e rite ana o ngā kēhi e rua o te adenocarcinoma wāhanga I i mua i te tangohanga o te pukahukahu. I whakaatuhia e te kēhi 3 he pona he 8 mm te diameter i te pito matau o runga me te tangohanga pleural. I whakaatuhia e te kēhi 4 he pona karāhe whenua totoka wāhanga e 9 mm te rahi i te pito maui o runga. Ko te tae hematoxylin me te eosin (H&E) o te kiko pukahukahu kua tangohia (pae tauine = 50 μm) e whakaatu ana i te tauira tipu acinar o te adenocarcinoma pukahukahu. E tohu ana ngā pere i ngā pona i kitea i runga i ngā whakaahua CT. Ko ngā whakaahua H&E he whakaahua tohu o ngā mara karukaru maha (>3) i tirotirohia e te tohunga mate.
Ina huihuia, e whakaatu ana ā mātou hua i te uara pea o ngā tohu koiora metabolite toto i roto i te tautuhi rerekētanga o ngā pona pūkahukahu, ka waiho pea tēnei hei wero i te aromatawai i te tirotiro CT.
I runga i tētahi papatohu matū rerekē kua whakamanahia, i whai mātou ki te tautuhi i ngā hononga koiora o ngā huringa nui o te pākia. I kitea e te tātari whakarei ara KEGG e MetaboAnalyst e 6 ngā ara kua tino rerekē i waenga i ngā rōpū e rua (LA vs. HC me LA vs. BN, kua whakatikatikaina te p ≤ 0.001, pānga > 0.01). I tohuhia ēnei huringa e ngā raruraru i roto i te pākia o te pyruvate, te pākia o te tryptophan, te pākia o te niacin me te nicotinamide, te glycolysis, te huringa TCA, me te pākia o te purine (Pikitia 5a). Kātahi ka whakahaerehia e mātou he metabolomics kua tohua hei manatoko i ngā huringa nui mā te whakamahi i te ine tino. Te whakatau i ngā matū noa i roto i ngā ara kua tino rerekē mā te triple quadrupole mass spectrometry (QQQ) mā te whakamahi i ngā paerewa matū pono. Kei roto i te Ripanga Tāpiri 4 ngā āhuatanga taupori o te tauira whāinga o te rangahau metabolomics. E ai ki ā mātou hua metabolomics ā-ao, i whakaū te tātaritanga tauanga kua piki ake te hypoxanthine me te xanthine, te pyruvate, me te lactate i roto i te LA ki te whakataurite ki te BN me te HC (Pikitia 5b, c, p <0.05). Heoi, kāore i kitea he rerekētanga nui i roto i ēnei metabolites i waenga i te BN me te HC.
Tātaritanga whakarei ake i te ara KEGG o ngā metabolites rerekē i roto i te rōpū LA ki te whakataurite ki ngā rōpū BN me HC. I whakamahia he Globaltest e rua-hiku, ā, i whakatikatikaina ngā uara p mā te whakamahi i te tikanga Holm-Bonferroni (p ≤ 0.001 i whakatikatikaina me te rahi o te pānga > 0.01). b–d Ngā kauwhata Violin e whakaatu ana i ngā taumata hypoxanthine, xanthine, lactate, pyruvate, me te tryptophan i roto i te serum HC, BN, me te LA i whakatauhia e LC-MS/MS (n = 70 mō ia rōpū). Ko ngā rārangi iraira mā me te pango e tohu ana i te waenga me te hauwhā, ia. e Kauwhata Violin e whakaatu ana i te whakaaturanga mRNA Log2TPM (tuhinga mō ia miriona) kua whakatauritea o SLC7A5 me QPRT i roto i te adenocarcinoma pūkahukahu (n = 513) ki te whakataurite ki te kiko pūkahukahu noa (n = 59) i roto i te huinga raraunga LUAD-TCGA. Ko te pouaka mā e tohu ana i te awhe interquartile, ko te rārangi pango whakapae i waenganui e tohu ana i te waenga, ā, ko te rārangi pango poutū e toro atu ana i te pouaka e tohu ana i te āputa whakawhirinaki 95% (CI). f Ko te kauwhata hononga Pearson o te whakaaturanga SLC7A5 me te GAPDH i roto i te adenocarcinoma pūkahukahu (n = 513) me te kiko pūkahukahu noa (n = 59) i roto i te huinga raraunga TCGA. Ko te rohe hina e tohu ana i te 95% CI. r, te tauwehenga hononga Pearson. g Ngā taumata tryptophan pūtau kua whakatauritea i roto i ngā pūtau A549 i whakawhitihia ki te mana whakahaere shRNA kore-motuhake (NC) me te shSLC7A5 (Sh1, Sh2) i whakatauhia e te LC-MS/MS. Ka whakaaturia te tātaritanga tatauranga o ngā tauira motuhake koiora e rima i roto i ia rōpū. h Ngā taumata pūtau o te NADt (NAD katoa, tae atu ki te NAD+ me te NADH) i roto i ngā pūtau A549 (NC) me ngā pūtau A549 knockdown SLC7A5 (Sh1, Sh2). Ka whakaaturia te tātaritanga tatauranga o ngā tauira motuhake koiora e toru i roto i ia rōpū. i I inehia te ngohe glycolytic o ngā pūtau A549 i mua atu, ā, i muri mai i te SLC7A5 knockdown mā te tere waikawa o waho o te pūtau (ECAR) (n = 4 ngā tauira motuhake koiora mō ia rōpū). 2-DG,2-deoxy-D-glucose. I whakamahia te whakamātautau t a te Tauira hiku-rua i roto i te (b–h). I roto i te (g–i), ko ngā pae hapa e tohu ana i te toharite ± SD, i mahia takitahi ia whakamātautau e toru ngā wā, ā, he rite tonu ngā hua. Ka whakaratohia ngā raraunga pūtake i roto i te ahua o ngā kōnae raraunga pūtake.
I runga i te pānga nui o te whakarerekētanga o te pākia o te tryptophan i roto i te rōpū LA, i arotakehia hoki e mātou ngā taumata tryptophan toto i roto i ngā rōpū HC, BN, me LA mā te whakamahi i te QQQ. I kitea e mātou kua heke te tryptophan toto i roto i te LA ki te whakataurite ki te HC, ki te BN rānei (p < 0.001, Pikitia 5d), he rite tonu ki ngā kitenga o mua ko ngā taumata tryptophan e rere ana i roto i te toto he iti ake i roto i ngā tūroro mate pukupuku pūkahukahu i ngā mana hauora mai i te rōpū whakahaere33,34,35. I kitea e tētahi atu rangahau e whakamahi ana i te PET/CT tracer 11C-methyl-L-tryptophan kua piki ake te wā pupuri tohu tryptophan i roto i te kiko mate pukupuku pūkahukahu ki te whakataurite ki ngā whara pai, ki te kiko noa rānei36. E whakapae ana mātou ko te hekenga o te tryptophan i roto i te toto LA ka whakaata pea i te tango kaha o te tryptophan e ngā pūtau mate pukupuku pūkahukahu.
E mōhiotia ana hoki ko te hua whakamutunga o te ara kynurenine o te tryptophan catabolism ko NAD+37,38, he matū nui mō te tauhohenga o te glyceraldehyde-3-phosphate me te 1,3-bisphosphoglycerate i roto i te glycolysis39. Ahakoa i arotahi ngā rangahau o mua ki te mahi a te tryptophan catabolism i roto i te whakahaere ārai mate, i whai mātou ki te whakamārama i te taunekeneke i waenga i te dysregulation tryptophan me ngā ara glycolytic i kitea i roto i te rangahau o nāianei. Ko te whānau kawe wairewa 7 mema 5 (SLC7A5) e mōhiotia ana he kawe tryptophan43,44,45. Ko te quinolinic acid phosphoribosyltransferase (QPRT) he whākōkī kei raro iho i te ara kynurenine e huri ana i te waikawa quinolinic ki NAMN46. I te tirotiro i te huinga raraunga LUAD TCGA i whakaatu ko te SLC7A5 me te QPRT i piki ake i roto i te kiko pukupuku i whakaritea ki te kiko noa (Pikitia 5e). I kitea tēnei pikinga i ngā wāhanga I me te II me ngā wāhanga III me te IV o te adenocarcinoma o te pūkahukahu (Pikitia Tāpiri 11), e tohu ana i ngā raruraru tōmua o te pākia o te tryptophan e pā ana ki te tumorigenesis.
I tua atu, i whakaatuhia e te huinga raraunga LUAD-TCGA he hononga pai i waenga i te whakaaturanga mRNA SLC7A5 me te GAPDH i roto i ngā tauira tūroro mate pukupuku (r = 0.45, p = 1.55E-26, Pikitia 5f). He rerekē, kāore i kitea he hononga nui i waenga i aua waitohu ira i roto i te kiko pūkahukahu noa (r = 0.25, p = 0.06, Pikitia 5f). Ko te hinganga o SLC7A5 (Pikitia Tāpiri 12) i roto i ngā pūtau A549 i tino whakaiti i ngā taumata tryptophan pūtau me te NAD(H) (Pikitia 5g,h), ka hua ake he ngoikore o te mahi glycolytic e inehia ana e te tere waikawa o waho (ECAR) (Pikitia 1). 5i). Nō reira, i runga i ngā huringa metabolic i roto i te serum me te kitenga in vitro, e whakapae ana mātou ka taea e te metabolism tryptophan te whakaputa NAD+ mā te ara kynurenine me te whai wāhi nui ki te whakatairanga i te glycolysis i roto i te mate pukupuku pūkahukahu.
Kua whakaatuhia e ngā rangahau he nui ngā pona pūkahukahu kāore i te tino kitea i kitea e te LDCT ka arahi pea ki te hiahia mō ētahi atu whakamātautau pēnei i te PET-CT, te koiora pūkahukahu, me te maimoatanga nui rawa atu nā te mea he tātaritanga teka-pai o te mate pukupuku.31 E ai ki te whakaaturanga i te Pikitia 6, i tautuhia e tā mātou rangahau he rōpū o ngā matū matū toto me te uara tātaritanga pea ka whakapai ake i te wehewehenga mōrearea me te whakahaere i muri mai o ngā pona pūkahukahu i kitea e te CT.
Ka arotakehia ngā pona pūkahukahu mā te whakamahi i te tomography rorohiko iti-horopeta (LDCT) me ngā āhuatanga whakaahua e tohu ana i ngā take pai, kino rānei. Ko te mutunga kore o ngā pona ka arahi pea ki ngā haerenga tirotiro auau, ngā wawaotanga koretake, me te maimoatanga nui rawa. Mā te whakaurunga o ngā whakarōpūtanga matūkai toto me te uara tātaritanga ka pai ake te aromatawai mōrearea me te whakahaere i muri mai o ngā pona pūkahukahu. Tomography tukunga positron PET.
E whakaatu ana ngā raraunga mai i te rangahau NLST o Amerika me te rangahau NELSON o Ūropi mā te tirotiro i ngā rōpū mōrearea nui mā te whakamahi i te tomography rorohiko iti-horopeta (LDCT) ka whakaiti i te mate pukupuku pūkahukahu1,3. Heoi, ko te aromatawai mōrearea me te whakahaere haumanu i muri mai o te maha o ngā pona pūkahukahu tūpono i kitea e te LDCT te mea tino uaua tonu. Ko te whāinga matua ko te arotau i te whakarōpūtanga tika o ngā kawa LDCT o nāianei mā te whakauru i ngā tohu koiora pono.
Kua tautuhia ētahi tohu koiora ngota, pērā i ngā matūkai toto, mā te whakataurite i te mate pukupuku pūkahukahu me ngā mana hauora15,17. I roto i tēnei rangahau, i arotahi mātou ki te whakamahinga o te tātaritanga matūkai toto hei wehewehe i waenga i ngā pona pūkahukahu pai me ngā pona kino i kitea ohoreretia e te LDCT. I whakatauritea e mātou te matūkai toto ā-ao o te mana hauora (HC), ngā pona pūkahukahu pai (BN), me ngā tauira adenocarcinoma pūkahukahu wāhanga I (LA) mā te whakamahi i te tātaritanga UPLC-HRMS. I kitea e mātou he rite ngā kōtaha matūkai o te HC me te BN, engari i whakaatuhia e te LA ngā huringa nui ki te whakataurite ki te HC me te BN. I tautuhia e mātou e rua ngā huinga matūkai toto e wehewehe ana i te LA mai i te HC me te BN.
Ko te kaupapa tautuhi LDCT o nāianei mō ngā pona pai me ngā pona kino e hangai ana ki te rahi, te kiato, te āhua me te tere tipu o ngā pona i roto i te wā30. Kua whakaatuhia e ngā rangahau o mua he hononga tata te rahi o ngā pona ki te tūponotanga o te mate pukupuku pūkahukahu. Ahakoa i roto i ngā tūroro mōrearea nui, ko te mōrearea o te mate pukupuku i roto i ngā pona <6 mm he <1%. Ko te mōrearea o te mate pukupuku mō ngā pona e ine ana i te 6 ki te 20 mm kei waenga i te 8% ki te 64%. Nō reira, e taunaki ana te Fleischner Society i te diameter tapahi o te 6 mm mō te whai i te CT auau.29 Heoi, kāore anō kia tino tika te aromatawai mōrearea me te whakahaere i ngā pona pūkahukahu kore-taurite (IPN) nui atu i te 6 mm 31. Ko te whakahaere o nāianei mō te mate ngakau whānau e hangai ana ki te tatari mataara me te aroturuki CT auau.
I runga i te metabolome kua whakamanahia, i whakaaturia e mātou mō te wā tuatahi te taupatupatu o ngā waitohu metabolomic i waenga i ngā tāngata hauora me ngā pona pai <6 mm. He rite te ōritetanga koiora ki ngā kitenga CT o mua, arā, ko te mōrearea o te mate pukupuku mō ngā pona <6 mm he iti iho i ngā tāngata kāore he pona.30 Me mōhio hoki ko ā mātou hua e whakaatu ana he nui te ōritetanga o ngā pona pai <6 mm me te ≥6 mm i roto i ngā kōtaha metabolomic, e kī ana he rite tonu te whakamāramatanga mahi o te take pai ahakoa te rahi o te pona. Nō reira, ka taea e ngā panera metabolite serum tātaritanga hou te whakarato i tētahi whakamātautau kotahi hei whakamātautau whakakore i te wā e kitea tuatahitia ana ngā pona i runga i te CT, ā, ka whakaiti pea i te aroturukitanga raupapa. I taua wā anō, i wehewehe te panera kotahi o ngā tohu koiora metabolic i ngā pona kino ≥6 mm te rahi mai i ngā pona pai, ā, i whakarato i ngā matapae tika mō ngā IPN o te rahi rite me ngā āhuatanga āhua koretake i runga i ngā whakaahua CT. I pai te mahi a tēnei whakarōpūtanga metabolism serum ki te matapae i te mate pukupuku o ngā pona ≥6 mm me te AUC o te 0.927. Ina huihuia, e whakaatu ana ā mātou hua ko ngā tohu motuhake o te pākia o te toto ka whakaata i ngā huringa pākia o te tinana i puta mai i te pukupuku, ā, he uara pea hei tohu mōrearea, ahakoa te rahi o te pona.
He mea nui, ko te adenocarcinoma o te pūkahukahu (LUAD) me te squamous cell carcinoma (LUSC) ngā momo matua o te mate pukupuku pūkahukahu kore-pūtau iti (NSCLC). I te mea he hononga kaha a LUSC ki te whakamahi tupeka47, ā, ko te LUAD te histology tino noa o ngā pona pūkahukahu tūpono i kitea i te tirotiro CT48, i hangaia motuhaketia tā mātou tauira whakarōpū mō ngā tauira adenocarcinoma wāhanga I. I arotahi hoki a Wang mā ki te LUAD, ā, i tautuhia e iwa ngā waitohu lipid mā te whakamahi i te lipidomics hei wehewehe i te mate pukupuku pūkahukahu wāhanga tuatahi mai i ngā tāngata hauora17. I whakamatauria e mātou te tauira whakarōpū o nāianei i runga i te 16 ngā take o te LUSC wāhanga I me te 74 ngā pona pai, ā, i kitea he iti te tika o te matapae LUSC (AUC 0.776), e kī ana he rerekē pea ngā waitohu metabolomic o te LUAD me te LUSC. Ina hoki, kua whakaatuhia he rerekē te LUAD me te LUSC i roto i te etiology, te takenga mai o te koiora me ngā hapa ira49. Nō reira, me whakauru ētahi atu momo histology ki roto i ngā tauira whakangungu mō te kimi i te mate pukupuku pūkahukahu i runga i te taupori i roto i ngā kaupapa tirotiro.
I konei, i tautuhia e mātou ngā ara e ono e tino rerekē ana i roto i te adenocarcinoma o te pūkahukahu, ki te whakatauritea ki ngā mana hauora me ngā pona pai. Ko te Xanthine me te hypoxanthine he matūkai noa o te ara metabolic purine. E ai ki ā mātou hua, ko ngā takawaenga e pā ana ki te metabolism purine i tino piki ake i roto i te serum, i ngā kiko rānei o ngā tūroro me te adenocarcinoma o te pūkahukahu, ki te whakatauritea ki ngā mana hauora, ki ngā tūroro rānei i te wāhanga mua o te whakaekenga15,50. Ko te taumata xanthine me te hypoxanthine serum teitei pea e whakaata ana i te anabolism e hiahiatia ana e ngā pūtau matepukupuku e tere haere ana te tipu. Ko te korenga o te metabolism glucose he tohu rongonui o te metabolism matepukupuku51. I konei, i kite mātou i te pikinga nui o te pyruvate me te lactate i roto i te rōpū LA ki te whakatauritea ki te rōpū HC me te BN, e rite ana ki ngā pūrongo o mua mō ngā rerekētanga o te ara glycolytic i roto i ngā kōtaha metabolome serum o ngā tūroro matepukupuku pūkahukahu kore-pūtau iti (NSCLC) me ngā mana hauora. he rite tonu ngā hua52,53.
He mea nui, i kitea e mātou he hononga whakamuri i waenga i te pākia o te pyruvate me te tryptophan i roto i te toto o ngā adenocarcinomas pūkahukahu. I heke ngā taumata tryptophan toto i roto i te rōpū LA ki te whakaritea ki te rōpū HC, BN rānei. He mea whakamīharo, i kitea i roto i tētahi rangahau nui o mua e whakamahi ana i tētahi rōpū tirohanga he hononga ngā taumata iti o te tryptophan e rere ana ki te pikinga o te mōrearea o te mate pukupuku pūkahukahu 54. Ko te tryptophan he waikawa amino nui e whiwhi katoa ana tātou mai i te kai. Ka whakatau mātou ko te whakahekenga o te tryptophan toto i roto i te adenocarcinoma pūkahukahu ka whakaata pea i te whakahekenga tere o tēnei metabolite. E mōhiotia whānuitia ana ko te hua whakamutunga o te catabolism tryptophan mā te ara kynurenine te pūtake o te hanganga NAD+ hou. Nā te mea ka puta te NAD+ i te ara whakaora, kāore anō kia whakatauhia te hiranga o te NAD+ i roto i te pākia o te tryptophan i roto i te hauora me te mate46. I whakaatuhia e tā mātou tātaritanga o te pātengi raraunga TCGA he nui ake te pikinga o te whakaaturanga o te kawe tryptophan solute transporter 7A5 (SLC7A5) i roto i te adenocarcinoma o te pūkahukahu ki te whakataurite ki ngā mana whakahaere noa, ā, he hononga pai ki te whakaaturanga o te whākōkī glycolytic GAPDH. Ko ngā rangahau o mua i aro nui ki te mahi a te catabolism tryptophan ki te pehi i te urupare ārai mate pukupuku40,41,42. I konei ka whakaatuhia e mātou ko te aukati i te tango tryptophan mā te patu i te SLC7A5 i roto i ngā pūtau mate pukupuku pūkahukahu ka hua ake he hekenga o ngā taumata NAD pūtau me te whakaiti i te mahi glycolytic. Hei whakarāpopototanga, ka whakaratohia e tā mātou rangahau he pūtake koiora mō ngā huringa o te metabolism serum e pā ana ki te huringa kino o te adenocarcinoma o te pūkahukahu.
Ko ngā whakarerekētanga EGFR ngā whakarerekētanga tino kitea i roto i ngā tūroro me te NSCLC. I roto i tā mātou rangahau, i kitea e mātou he rite tonu ngā āhua metabolomic o ngā tūroro me te whakarerekētanga EGFR (n = 41) ki ngā tūroro me te EGFR momo-mohoao (n = 31), ahakoa i kitea e mātou he iti iho ngā taumata serum o ētahi tūroro whakarerekētanga EGFR i roto i ngā tūroro acylcarnitine. Ko te mahi kua whakapumautia o ngā acylcarnitine ko te kawe i ngā rōpū acyl mai i te cytoplasm ki roto i te matrix mitochondrial, ka arahina ki te waikura o ngā waikawa ngako hei whakaputa pūngao 55. E rite ana ki ā mātou kitenga, i kitea anō hoki e tētahi rangahau tata nei ngā āhua metabolome rite i waenga i ngā pukupuku whakarerekētanga EGFR me te momo-mohoao EGFR mā te tātari i te metabolome ā-ao o te 102 tauira kiko adenocarcinoma pūkahukahu 50. He mea whakamīharo, i kitea anō hoki te ihirangi acylcarnitine i roto i te rōpū whakarerekētanga EGFR. Nō reira, mehemea e whakaata ana ngā huringa o ngā taumata acylcarnitine i ngā huringa metabolic i puta mai i te EGFR me ngā ara ngota ngota kei raro, tera pea he mea tika kia rangahauhia anō.
Hei whakamutunga, ka whakatūhia e tā mātou rangahau he whakarōpūtanga pākia toto mō te tautuhi rerekētanga o ngā pona pūkahukahu, ā, ka whakaarohia he tukanga mahi ka taea te arotau i te aromatawai mōrearea me te whakahaere i te whakahaere haumanu i runga i te tirotiro CT scan.
I whakaaetia tēnei rangahau e te Komiti Matatika o te Hōhipera Mate Pukupuku o te Whare Wānanga o Sun Yat-sen, te Hōhipera Hononga Tuatahi o te Whare Wānanga o Sun Yat-sen, me te Komiti Matatika o te Hōhipera Mate Pukupuku o te Whare Wānanga o Zhengzhou. I roto i ngā rōpū kitenga me te whakamana ā-roto, 174 ngā toto mai i ngā tāngata hauora me 244 ngā toto mai i ngā pona pai i kohia mai i ngā tāngata e tirotirohia ana e te Tari Whakahaere me te Ārai Mate Pukupuku, te Pokapū Mate Pukupuku o te Whare Wānanga o Sun Yat-sen, me 166 ngā pona pai. toto. I kohia ngā adenocarcinoma pūkahukahu Wāhanga I mai i te Pokapū Mate Pukupuku o te Whare Wānanga o Sun Yat-sen. I roto i te rōpū whakamana ā-waho, e 48 ngā take o ngā pona pai, 39 ngā take o te adenocarcinoma pūkahukahu wāhanga I mai i te Hōhipera Hononga Tuatahi o te Whare Wānanga o Sun Yat-sen, me 24 ngā take o te adenocarcinoma pūkahukahu wāhanga I mai i te Hōhipera Mate Pukupuku o Zhengzhou. I kohia hoki e te Pokapū Mate Pukupuku o te Whare Wānanga o Sun Yat-sen e 16 ngā take o te mate pukupuku pūkahukahu pūtau squamous wāhanga I hei whakamātautau i te kaha tātaritanga o te whakarōpūtanga metabolic kua whakatūria (kei te whakaatuhia ngā āhuatanga o te tūroro i te Ripanga Tāpiri 5). I kohia ngā tauira mai i te rōpū kitenga me te rōpū whakamana ā-roto i waenganui i a Hanuere 2018 me Mei 2020. I kohia ngā tauira mō te rōpū whakamana ā-waho i waenganui i a Ākuhata 2021 me Oketopa 2022. Hei whakaiti i te whakaaro pākiki ira tangata, tata rite te maha o ngā tāne me ngā wāhine i tohaina ki ia rōpū. Te Rōpū Kitea me te Rōpū Arotake ā-Roto. I whakatauhia te ira tangata o te kaiuru i runga i te pūrongo whaiaro. I whiwhi whakaaetanga whai whakaaro mai i ngā kaiuru katoa, ā, kāore i tukuna he utu. Ko ngā kaupapa he pona pai ko te hunga he kaute CT scan pumau i te 2 ki te 5 tau i te wā o te tātaritanga, engari kotahi te take mai i te tauira whakamana ā-waho, i kohia i mua i te pokanga, ā, i tātaritatia mā te histopathology. Haunga te bronchitis mau tonu. I kohia ngā take adenocarcinoma pūkahukahu i mua i te tango i te pūkahukahu, ā, i whakaūtia mā te tātaritanga pathological. I kohia ngā tauira toto nohopuku i roto i ngā ngongo wehenga toto me te kore he anticoagulants. I whakapakeketia ngā tauira toto mō te 1 hāora i te pāmahana rūma, kātahi ka poipoia ki te 2851 × g mō te 10 meneti i te 4°C hei kohi i te supernatant toto. I whakatiohia ngā wāhanga toto i te -80°C tae noa ki te tangohanga o te matū matū. I kohia e te Tari Ārai Mate Pukupuku me te Whakamātautau Hauora o te Pokapū Mate Pukupuku o te Whare Wānanga o Sun Yat-sen he puna toto mai i te 100 o ngā kaituku hauora, tae atu ki te maha ōrite o ngā tāne me ngā wāhine i waenga i te 40 ki te 55 tau te pakeke. I whakaranua ngā rahinga ōrite o ia tauira kaituku, ā, i wehea te puna hua, ā, i rongoatia ki te -80°C. I whakamahia te ranunga toto hei rauemi tohutoro mō te whakahaere kounga me te paerewa raraunga.
I whakarewaina te toto tohutoro me ngā tauira whakamātautau, ā, i tangohia ngā matū matū ma te whakamahi i te tikanga tango whakakotahi (MTBE/methanol/wai) 56. I roto i te poto, i konatunatua te 50 μl o te toto ki te 225 μl o te methanol makariri me te 750 μl o te methyl tert-butyl ether makariri (MTBE). Whakaohokia te ranunga ka whaowhia ki runga i te hukapapa mō te 1 hāora. Kātahi ka konatunatua ngā tauira, ā, ka konatunatua ki te vortex ki te 188 μl o te wai MS-grade kei roto ngā paerewa ā-roto (13C-lactate, 13C3-pyruvate, 13C-methionine, me te 13C6-isoleucine, i hokona mai i Cambridge Isotope Laboratories). Kātahi ka porotītahahia te ranunga ki te 15,000 × g mō te 10 meneti i te 4 °C, ā, ka whakawhitia te wāhanga o raro ki roto i ngā ngongo e rua (125 μL ia ngongo) mō te tātari LC-MS i ngā aratau pai me te aratau kino. Hei whakamutunga, i mimitia te tauira kia maroke i roto i te mīhini whakaemi korehau tere-teitei.
I hangaia anō ngā matū matū maroke ki roto i te 120 μl o te 80% acetonitrile, i hurihia mō te 5 meneti, ā, i whakatōngia ki te 15,000 × g mō te 10 meneti i te 4°C. I whakawhitia ngā wai whakapūkara ki roto i ngā ipu karāhe amber me ngā whakaurunga iti mō ngā rangahau metabolomics. Tātari metabolomics kāore i te whaingahia i runga i te tūāpapa ultra-performance liquid chromatography-high-resolution mass spectrometry (UPLC-HRMS). I wehea ngā matū matū matū mā te whakamahi i te pūnaha Dionex Ultimate 3000 UPLC me te pou ACQUITY BEH Amide (2.1 × 100 mm, 1.7 μm, Waters). I roto i te aratau katote pai, ko ngā āhua nekeneke he 95% (A) me te 50% acetonitrile (B), kei roto i ia mea he 10 mmol/L ammonium acetate me te 0.1% waikawa formic. I te aratau kino, i roto i ngā wāhanga neke A me B he 95% me te 50% acetonitrile, ia, i roto i ngā wāhanga e rua he 10 mmol/L ammonium acetate, pH = 9. Ko te kaupapa pikinga penei: 0–0.5 meneti, 2% B; 0.5–12 meneti, 2–50% B; 12–14 meneti, 50–98% B; 14–16 meneti, 98% B; 16–16.1 meneti, 98 –2% B; 16.1–20 meneti, 2% B. I puritia te pou i te 40°C me te tauira i te 10°C i roto i te autosampler. Ko te tere rere he 0.3 ml/min, ko te rōrahi werohanga he 3 μl. I whakahaerehia he ine matawai papatipu Q-Exactive Orbitrap (Thermo Fisher Scientific) me te pūtake electrospray ionization (ESI) i roto i te aratau matawai katoa, ā, i honoa ki te aratau aroturuki ddMS2 hei kohikohi i ngā rahinga raraunga nui. I whakatakotoria ngā tawhā MS penei: ngaohiko rehu +3.8 kV/- 3.2 kV, pāmahana capillary 320°C, hau tiaki 40 arb, hau āwhina 10 arb, pāmahana whakamahana probe 350°C, whānuitanga matawai 70–1050 m/h, taumira. 70 000. I kohia ngā raraunga mā te whakamahi i te Xcalibur 4.1 (Thermo Fisher Scientific).
Hei aromatawai i te kounga o ngā raraunga, i hangaia ngā tauira whakahaere kounga (QC) mā te tango i te 10 μL o ngā wāhanga o te wai i roto i ia tauira. E ono ngā werohanga tauira whakahaere kounga i tātarihia i te tīmatanga o te raupapa tātari hei aromatawai i te pumau o te pūnaha UPLC-MS. Kātahi ka whakaurua ngā tauira whakahaere kounga ki roto i te puranga i ia wā. I tātarihia ngā puranga 11 katoa o ngā tauira toto i roto i tēnei rangahau mā te LC-MS. I whakamahia ngā wāhanga o te ranunga puna toto mai i te 100 o ngā kaituku hauora hei rauemi tohutoro i roto i ngā puranga hei aroturuki i te tukanga tango me te whakarite mō ngā pānga puranga-ki-puranga. I whakahaerehia te tātari metabolomics kore-whaiaro o te rōpū kitenga, te rōpū whakamana ā-roto, me te rōpū whakamana ā-waho i te Pokapū Metabolomics o Te Whare Wānanga o Sun Yat-sen. I tātarihia hoki e te taiwhanga ā-waho o te Pokapū Tātari me te Whakamātautau o Te Whare Wānanga o Guangdong Technology te 40 tauira mai i te rōpū ā-waho hei whakamātautau i te mahi a te tauira whakarōpū.
I muri i te tangohanga me te hanganga anō, i inehia te rahinga tino o ngā matū matū i roto i te toto mā te whakamahi i te ultra-high performance liquid chromatography-tandem mass spectrometry (Agilent 6495 triple quadrupole) me te pūtake electrospray ionization (ESI) i roto i te aratau aroturuki tauhohenga maha (MRM). I whakamahia he pou ACQUITY BEH Amide (2.1 × 100 mm, 1.7 μm, Waters) hei wehe i ngā matū matū. Ko te wāhanga nekeneke i titoa mai i te 90% (A) me te 5% acetonitrile (B) me te 10 mmol/L ammonium acetate me te 0.1% otinga ammonia. Ko te kaupapa gradient i penei: 0–1.5 min, 0% B; 1.5–6.5 min, 0–15% B; 6.5–8 min, 15% B; 8–8.5 min, 15%–0% B; 8.5–11.5 min, 0%B. I puritia te pou i te 40 °C, ā, i te 10 °C te tauira i roto i te autosampler. Ko te tere rere he 0.3 mL/min, ā, ko te rōrahi werohanga he 1 μL. Ko ngā tawhā MS i whakatakotoria penei: te ngaohiko capillary ±3.5 kV, te pēhanga nebulizer 35 psi, te rere hau o te uhi 12 L/min, te pāmahana hau o te uhi 350°C, te pāmahana hau whakamaroke 250°C, me te rere hau whakamaroke 14 l/min. Ko ngā huringa MRM o te tryptophan, pyruvate, lactate, hypoxanthine me te xanthine he 205.0–187.9, 87.0–43.4, 89.0–43.3, 135.0–92.3 me te 151.0–107.9 ia. I kohia ngā raraunga mā te whakamahi i te Mass Hunter B.07.00 (Agilent Technologies). Mō ngā tauira toto, i inehia te tryptophan, te pyruvate, te lactate, te hypoxanthine, me te xanthine mā te whakamahi i ngā ānau whakatikatika o ngā otinga ranunga paerewa. Mō ngā tauira pūtau, i whakatauritehia te nui o te tryptophan ki te paerewa ā-roto me te papatipu pūmua pūtau.
I whakahaerehia te tango tihi (m/z me te wā pupuri (RT)) mā te whakamahi i te Compound Discovery 3.1 me te TraceFinder 4.0 (Thermo Fisher Scientific). Hei whakakore i ngā rerekētanga pea i waenga i ngā puranga, i wehea ia tihi āhuatanga o te tauira whakamātautau e te tihi āhuatanga o te rauemi tohutoro mai i te puranga kotahi hei whiwhi i te nui whanaunga. Ko ngā paerewa rerekētanga whanaunga o ngā paerewa ā-roto i mua me muri i te whakapūmautanga e whakaaturia ana i roto i te Ripanga Tāpiri 6. Ko ngā rerekētanga i waenga i ngā rōpū e rua i tohuhia e te tere kitenga teka (FDR<0.05, whakamātautau rarangi hainatia a Wilcoxon) me te huringa kopa (>1.2, <0.83 rānei). Ko ngā raraunga MS mata o ngā āhuatanga i tangohia me ngā raraunga MS i whakatikatikaina i te toto tohutoro e whakaaturia ana i roto i te Raraunga Tāpiri 1 me te Raraunga Tāpiri 2. I whakahaerehia te tuhipoka tihi i runga i ngā taumata tautuhi e whā kua tautuhia, tae atu ki ngā metabolites kua tautuhia, ngā matū kua tohua, ngā akomanga matū kua tohua, me ngā matū kāore i te mōhiotia 22. I runga i ngā rapunga pātengi raraunga i roto i te Compound Discovery 3.1 (mzCloud, HMDB, Chemspider), i whiriwhiria ngā matū koiora me ngā paerewa whaimana MS/MS, ngā tuhipoka ōrite rānei i roto i te mzCloud (kaute > 85) Chemspider rānei hei takawaenga i waenga i te metabolome rerekē. Kei roto i te Raraunga Tāpiri 3 ngā tuhipoka tihi mō ia āhuatanga. I whakamahia te MetaboAnalyst 5.0 mō te tātari kotahi-taurangi o te nui o te metabolite kua whakatauritea. I arotakehia hoki e te MetaboAnalyst 5.0 te tātari whakarei ara KEGG i runga i ngā metabolite rerekē. I tātarihia te tātari wāhanga matua (PCA) me te tātari wehewehe tapawhā iti rawa wāhanga (PLS-DA) mā te whakamahi i te mōkihi pūmanawa ropls (v.1.26.4) me te whakataurite puranga me te tauine aunoa. I hangaia te tauira biomarker metabolite tino pai mō te matapae i te mate pukupuku pona mā te whakamahi i te whakatauira logistic rua me te iti rawa o te whakahekenga tino me te kaiwhakahaere kōwhiringa (LASSO, mōkihi R v.4.1-3). I whakaahuahia te mahi a te tauira wehewehe i roto i ngā huinga kimi me te whakamana mā te whakatau tata i te AUC i runga i te tātaritanga ROC e ai ki te mōkihi pROC (v.1.18.0.). I whiwhihia te tapahi tūponotanga tino pai i runga i te taupū Youden mōrahi o te tauira (aronga + motuhaketanga – 1). Ko ngā tauira he iti iho, he nui ake rānei i te paepae ka matapaetia he pona pai me te adenocarcinoma pūkahukahu.
I whakatipuria ngā pūtau A549 (#CCL-185, American Type Culture Collection) i roto i te reo F-12K kei roto ko te 10% FBS. I whakaurua ngā raupapa RNA makawe poto (shRNA) e aro ana ki a SLC7A5 me te mana whakahaere kore-aronga (NC) ki roto i te weriweri lentiviral pLKO.1-puro. Ko ngā raupapa antisense o shSLC7A5 e whai ake nei: Sh1 (5′-GGAGAAACCTGAATGAACAGTT-3′), Sh2 (5′-GCCGTGGACTTCGGGAACTAT-3′). I hokona mai i Cell Signaling Technology ngā paturopi ki a SLC7A5 (#5347) me te tubulin (#2148). I whakamahia ngā paturopi ki a SLC7A5 me te tubulin i te waimehatanga o te 1:1000 mō te tātaritanga Western blot.
Ka ine te Whakamātautau Taumahatanga Glycolytic Seahorse XF i ngā taumata waikawa o waho o te pūtau (ECAR). I roto i te whakamātautau, i tukuna te hukahuka, te oligomycin A, me te 2-DG i ia wā hei whakamātautau i te kaha glycolytic pūtau e inehia ana e te ECAR.
Ko ngā pūtau A549 i whakawhitihia ki te mana whakahaere kore-whāinga (NC) me te shSLC7A5 (Sh1, Sh2) i pania mō te pō ki roto i ngā rihi 10 cm te whānui. I tangohia ngā matū matū pūtau ki te 1 ml o te methanol wai makariri 80%. I waruhia ngā pūtau i roto i te otinga methanol, i kohia ki roto i tētahi ngongo hou, ā, i whakatōngia ki te 15,000 × g mō te 15 meneti i te 4°C. Kohia he 800 µl o te wai i roto i te ipu, ā, whakamarokehia mā te whakamahi i tētahi mīhini whakapūmau korehau tere-teitei. Kātahi ka tātarihia ngā pire matū matū maroke mō ngā taumata tryptophan mā te whakamahi i te LC-MS/MS e ai ki te whakaahuatanga i runga ake nei. I inehia ngā taumata NAD(H) pūtau i roto i ngā pūtau A549 (NC me te shSLC7A5) mā te whakamahi i tētahi kete tae NAD+/NADH tauanga (#K337, BioVision) e ai ki ngā tohutohu a te kaihanga. I inehia ngā taumata pūmua mō ia tauira hei whakataurite i te nui o ngā matū matū.
Kāore i whakamahia he tikanga tatauranga hei whakatau i te rahi o te tauira i te tīmatanga. Ko ngā rangahau metabolomics o mua i aro ki te kimi i ngā tohu koiora15,18 kua whakaarohia hei paearu mō te whakatau i te rahi, ā, ki te whakatauritea ki ēnei pūrongo, he rawaka tā mātou tauira. Kāore i tangohia ngā tauira mai i te rōpū rangahau. I tohaina matapōkeretia ngā tauira toto ki tētahi rōpū kitenga (306 ngā take, 74.6%) me tētahi rōpū whakamana ā-roto (104 ngā take, 25.4%) mō ngā rangahau metabolomics kāore i te arotahi. I tīpakohia matapōkeretia hoki e mātou he 70 ngā take mai i ia rōpū mai i te huinga kitenga mō ngā rangahau metabolomics kua arotahi. I matapōkeretia ngā kairangahau ki te tohatoha rōpū i te wā o te kohikohinga me te tātari raraunga LC-MS. Kei roto i ngā wāhanga Hua, Pikitia Pūrākau, me ngā Tikanga ngā tātari tatauranga o ngā raraunga metabolomics me ngā whakamātautau pūtau. E toru ngā wā i mahia motuhake ai te ine i te tryptophan pūtau, NADT, me te mahi glycolytic me ngā hua ōrite.
Mō ētahi atu kōrero e pā ana ki te hoahoa o te rangahau, tirohia te Whakarāpopototanga o te Pūrongo Pūkete Taiao e pā ana ki tēnei tuhinga.
Ko ngā raraunga MS mata o ngā āhuatanga i tangohia me ngā raraunga MS kua whakatauritea o te serum tohutoro e whakaaturia ana i roto i te Raraunga Tāpiri 1 me te Raraunga Tāpiri 2. Ko ngā tuhipoka tihi mō ngā āhuatanga rerekētanga e whakaaturia ana i roto i te Raraunga Tāpiri 3. Ka taea te tango i te huinga raraunga LUAD TCGA mai i https://portal.gdc.cancer.gov/. Kei roto i te raraunga pūtake ngā raraunga whakauru mō te tuhi i te kauwhata. Kei roto i tēnei tuhinga ngā raraunga pūtake.
Rōpū Rangahau Tirotiro Pūkahukahu ā-Motu, me ētahi atu. Te whakaiti i te mate pukupuku pūkahukahu mā te whakamahi i te tomography rorohiko iti-horopeta. Te Tai Tokerau o Ingarangi. J. Med. 365, 395–409 (2011).
Kramer, BS, Berg, KD, Aberle, DR me Prophet, PC Te tirotiro i te mate pukupuku pūkahukahu mā te whakamahi i te CT helical iti-horopeta: ngā hua mai i te National Lung Screening Study (NLST). J. Med. Screen 18, 109–111 (2011).
De Koning, HJ, et al. Te whakaiti i te mate pukupuku pūkahukahu mā te tirotiro CT rahi i roto i tētahi whakamātautau matapōkere. Te Tai Tokerau o Ingarangi. J. Med. 382, 503–513 (2020).
Wā tuku: Hepetema-18-2023