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《CIK逆轉(zhuǎn)耐順鉑人肺腺癌細(xì)胞對順鉑耐藥與ERCC1的相關(guān)性-論文.pdf》由會(huì)員上傳分享，免費(fèi)在線閱讀，更多相關(guān)內(nèi)容在應(yīng)用文檔-天天文庫。
1、第23071卷4年第6月3期遵義醫(yī)學(xué)院學(xué)報(bào)V01．37No．3JournalofZunyiMedicalUniversityJun．2014CIK逆轉(zhuǎn)耐順鉑人肺腺癌細(xì)胞對順鉑耐藥與ERCC1的相關(guān)性田昕，李寧，李冉(遵義醫(yī)學(xué)院附屬腫瘤醫(yī)院胸部腫瘤科，貴州遵義563099)[摘要]目的觀察臍血來源的CIK細(xì)胞對A549／DDP細(xì)胞的耐藥逆轉(zhuǎn)作用。并探討CIK逆轉(zhuǎn)耐藥機(jī)制是否與核苷酸切除修復(fù)交叉互補(bǔ)基因1(excisionrepaircrosscomplementationgroup1，ERCC1)的
2、表達(dá)存在相關(guān)性。方法采集臍血單個(gè)核細(xì)胞經(jīng)細(xì)胞因子體外誘導(dǎo)獲得CIK細(xì)胞(cytokineinducedkillerceils，CIK)，MIrr法檢測順鉑(cisplatin，DDP)對A549細(xì)胞和A549／DDP細(xì)胞的體外殺傷作用，以及CIK細(xì)胞與A549／DDP細(xì)胞共培養(yǎng)前后對DDP敏感性的變化。Real—timeRT—PCR法檢測A549細(xì)胞、A549／DDP細(xì)胞以及CIK干預(yù)后的A549／DDP細(xì)胞中ERCC1mRNA的表達(dá)情況。Westernblot法檢測A549細(xì)胞、A549／DDP
3、細(xì)胞以及CIK干預(yù)后的A549／DDP細(xì)胞中ERCC1蛋白的表達(dá)情況。結(jié)果A549／DDP細(xì)胞對DDP耐藥，耐藥指數(shù)為2．391。CIK細(xì)胞能逆轉(zhuǎn)A549／DDP細(xì)胞對DDP的耐藥性，逆轉(zhuǎn)倍數(shù)為3．967。Real—timeRT—PCR、West—ernblot測得A549／DDP細(xì)胞中ERCC1mRNA和蛋白表達(dá)量比A549細(xì)胞高(P<0．05)，CIK細(xì)胞共培養(yǎng)前后A549／DDP細(xì)胞中ERCC1mRNA和蛋白表達(dá)量無明顯變化。結(jié)論CIK細(xì)胞能逆轉(zhuǎn)A549／DDP細(xì)胞對DDP的耐藥性，其逆轉(zhuǎn)耐
4、藥的機(jī)制與ERCC1的表達(dá)無明顯相關(guān)性。[關(guān)鍵詞]肺癌；A549細(xì)胞；順鉑；耐藥逆轉(zhuǎn)；CIK細(xì)胞[中圖法分類號]R734．2[文獻(xiàn)標(biāo)志碼]A[文章編號]1000—2715(2014)03-0338-05StudyonthecorrelationbetweenERCC1andthemerchanismsunderlyingcy·tokineinducedkillercells——reversedcisplatinresistanceinA549／DDPCellsTianXin，LNing，LiRan
5、(DepartmentofThoracic，TheAffiliatedHospitalofZunyiMedicalUniversity，ZunyiGuizhou563099，China)[Abstract]ObjectiveToinvestigatetheeffectsofcordblood—derivedcytokine—inducedkiller(CIK)cellsonreversingcisplatinresistanceintheDDP——resistanthumanlungadenoca
6、rcinomacelllineA549／DDPandobservewhethertherewascorrelationbetweentheseeffectsandexcisionrepaircrosscomplementationgroup1(ERCC1)expression．MethodsThesingleumbilicalcordbloodmononuclearcellwasinducedbycytokineinvitrotobe—comeCIKcells．MTFassaywasapplied
7、toevaluatethecytotoxicactivityofDDPagainstA549／DDPcellsandA549cellsinvitroandthesensitivityofA549／DDPcellsCIKtoDDPbeforeandaftertheco—culturewithCIK．ThemRNAandproteinexpressionsofERCC1inA549cells。A549／DDPcellsandA549／DDPco—culturedwithCIKcellsweredete
8、ctedbyreal—timeRT—PCRandwesternblotanalysis．ResultsTheA549／DDPcellswasresistanttoDDPwiththeresistanceindexof2．391．CIKcellsdecreaseddrugresistancetocisplatinwiththere．versalfoldof3．967．TheERCC1mRNAandproteinexprossionswerehigherinA549／DDPthanth
