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《CIK逆轉(zhuǎn)耐順鉑人肺腺癌細胞對順鉑耐藥與ERCC1的相關(guān)性-論文.pdf》由會員上傳分享,免費在線閱讀,更多相關(guān)內(nèi)容在應用文檔-天天文庫。
1、第23071卷4年第6月3期遵義醫(yī)學院學報V01.37No.3JournalofZunyiMedicalUniversityJun.2014CIK逆轉(zhuǎn)耐順鉑人肺腺癌細胞對順鉑耐藥與ERCC1的相關(guān)性田昕,李寧,李冉(遵義醫(yī)學院附屬腫瘤醫(yī)院胸部腫瘤科,貴州遵義563099)[摘要]目的觀察臍血來源的CIK細胞對A549/DDP細胞的耐藥逆轉(zhuǎn)作用。并探討CIK逆轉(zhuǎn)耐藥機制是否與核苷酸切除修復交叉互補基因1(excisionrepaircrosscomplementationgroup1,ERCC1)的
2、表達存在相關(guān)性。方法采集臍血單個核細胞經(jīng)細胞因子體外誘導獲得CIK細胞(cytokineinducedkillerceils,CIK),MIrr法檢測順鉑(cisplatin,DDP)對A549細胞和A549/DDP細胞的體外殺傷作用,以及CIK細胞與A549/DDP細胞共培養(yǎng)前后對DDP敏感性的變化。Real—timeRT—PCR法檢測A549細胞、A549/DDP細胞以及CIK干預后的A549/DDP細胞中ERCC1mRNA的表達情況。Westernblot法檢測A549細胞、A549/DDP
3、細胞以及CIK干預后的A549/DDP細胞中ERCC1蛋白的表達情況。結(jié)果A549/DDP細胞對DDP耐藥,耐藥指數(shù)為2.391。CIK細胞能逆轉(zhuǎn)A549/DDP細胞對DDP的耐藥性,逆轉(zhuǎn)倍數(shù)為3.967。Real—timeRT—PCR、West—ernblot測得A549/DDP細胞中ERCC1mRNA和蛋白表達量比A549細胞高(P<0.05),CIK細胞共培養(yǎng)前后A549/DDP細胞中ERCC1mRNA和蛋白表達量無明顯變化。結(jié)論CIK細胞能逆轉(zhuǎn)A549/DDP細胞對DDP的耐藥性,其逆轉(zhuǎn)耐
4、藥的機制與ERCC1的表達無明顯相關(guān)性。[關(guān)鍵詞]肺癌;A549細胞;順鉑;耐藥逆轉(zhuǎn);CIK細胞[中圖法分類號]R734.2[文獻標志碼]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