雷帕霉素对胃癌SGC7901细胞增殖影响及其机制探讨

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1、雷帕霉素对胃癌SGC7901细胞增殖影响及其机制探讨中文摘要目的研究雷帕霉素靶向抑制mTOR信号通路对胃癌SGC7901细胞增殖的影响及其可能机制，为胃癌的药物治疗提供新的思路和方法.方法体外培养胃癌SGC7901细胞，用免疫细胞化学法检测缺氧条件下胃癌SGC7901细胞中mTOR及HIF-1α蛋白表达情况。细胞经不同浓度雷帕霉素分别作用12h、24h、48h、72h，CCK-8法检测雷帕霉素对细胞增殖的抑制作用。半定量RT-PCR检测细胞mT0R和HIF-1αmRNA表达改变情况,WesternBlot法检测细胞mT0R和HIF-1α蛋白表达改变情况。实验所得数据采用SPSS

2、13.0统计分析软件进行处理。结果免疫细胞化学显示，缺氧条件下胃癌SGC7901细胞胞浆中mTOR呈阳性表达，细胞核和细胞浆中HIF-1α均呈阳性表达。雷帕霉素能显著抑制胃癌SGC7901细胞增殖，抑制率随药物浓度和作用时间增加而增加，呈时间-剂量依赖关系。雷帕霉素作用后，细胞mT0R和HIF-1αmRNA及蛋白表达水平均明显下调。结论缺氧条件下，胃癌SGC7901细胞存在mTOR/HIF-1α信号通路的异常活化。在体外雷帕霉素能显著抑制胃癌SGC7901细胞增殖。缺氧条件下，雷帕霉素抑制mTOR/HIF-1α信号通路是其可能机制之一。关键词雷帕霉素胃肿瘤哺乳动物雷帕霉素靶蛋白

3、缺氧诱导因子-1α药物治疗2EffectofRapamycinonhumangastriccarcinomaSGC7901cellsproliferationandapproachofthemechanismsABSTRACTObjectiveToinvestigatetheeffectsofRapamycinoncellproliferationinhumangastriccarcinomaSGC7901cellsandthepotentialmechanismsinvolvedintheinhibitoryeffectsofthemTORsignalingpathway.P

4、rovidethenewideasandoriginalstrategyaboutthedrugtreatmentforhumangastriccarcinoma.MethodsSGC7901cellswereculturedinvitro．TheexpressionofmTORandHIF-1αproteinswasdeterminedbyimmunocytochemicalmethodinSGC7901cellsculturedunderhypoxicconditions．DifferentconcentrationsofRapamycinwereincubatedwith

5、SGC7901cellsfor12hours、24hours、48hoursand72hours．CellproliferationwasmeasuredbyCCK-8assayinRapamycin—treatedSGC7901cells．TheexpressionlevelsofmTORandHIF—lαmRNAwasdetectedbysemi-quantitativeRT-PCR．ThechangeofmTORandHIF—lαproteinswasdeterminedbyWesternBlot.TheexperimentaldatawereprocessedbySPS

6、S13.0.ResultsImmunocytochemistryshowedthatunderhypoxicconditions,mTORwaspositivelyexpressedinthecytoplasmandthatHIF—lαinthenucleusandcytoplasmofSGC7901cells.RapamycincouldsignificantlyinhibittheproliferationofSGC7901cellsinaconcentration-dependentandtime-dependentmanner．Theexpressionlevelsof

7、mTOR、HIF—lαmRNAandmToR、HIF—lαproteinsweresignificantlylowerinRapamycin—treatedSGC7901cellsthanthoseinuntreatedcells．ConclusionsmTOR/HIF—lαsignalingpathwayisabnormallyactivatedinSGC7901cellsunderhypoxicconditions．Rapamycincouldsignificantlyinhibitth