摘要
目的探讨雷帕霉素(RPM)对体外培养的胰腺癌PC-2细胞增殖和凋亡的影响。方法以10～50 nmol/L不同浓度RPM处理PC-2细胞,四甲基偶氮唑盐法检测不同浓度、不同时间RPM处理后的PC-2细胞生长情况;AnnexinV-FITC/PI双标记染色,流式细胞术检测肿瘤细胞凋亡情况;反转录-聚合酶链反应方法检测mTOR mRNA表达。结果不同浓度的RPM作用于PC-2细胞0～96 h后,对PC-2细胞的生长有明显的抑制作用,呈时间和浓度依赖性;流式细胞仪检测结果显示,10～50 nmol/L RPM均能诱导不同程度的细胞凋亡,且10～30 nmol/L RPM组以早期凋亡细胞为主,随着RPM浓度的增高,晚期凋亡细胞明显增多;反转录-聚合酶链反应结果显示,与空白对照组相比,药物处理组mRNA相对表达量均有不同程度的抑制效应,而且随着RPM浓度增高,mTOR mRNA水平表达逐渐降低,RPM与mTOR mRNA表达间表现出较好的剂量依赖性关系(P<0.05)。结论 RPM可有效抑制胰腺癌PC-2细胞增殖,并诱导细胞凋亡,其作用机制可能与下调mTOR基因表达有关。
Objective To investigate the effects of Rapamycin（RPM）in suppressing proliferation and inducing apoptosis of pancreatic carcinoma PC-2 cells in vitro.Methods MTT assay was used to examine the proliferation of PC-2 cells treated by different concentration（10-50 nmol/L）RPM after 0-96 hours.Flow cytometry was performed to analyze the apoptosis of PC-2 cells by staining with annexin V-FITC/PI.The expression of mTOR was measured by semi-quantitive reverse transcription polymerase chain reaction（RT-PCR）method.Results The inhibition of proliferation of PC-2 cells in vitro was observerd in time and dose dependent.Flow cytometry assay also showed RPM had positive effect on apoptosis.Cells treated with 10-30 nmol/L RPM are mainly composed of early apoptotic cells.While,cells treated with high dose RPM are mainly composed of late apoptotic cells.RPM could effectively down-regulate the expression of mTOR in the mRNA levels.The relative expression of mTOR to RPM was increased in a dose-dependent manner（P〈0.05）.Conclusion RPM could effectively inhibit proliferation and induce apoptosis in PC-2 cells.The mechanism could be related to down-expression of mTOR.
出处
《医学综述》
2011年第18期2826-2829,共4页
Medical Recapitulate
基金
陕西省科技攻关项目(2010K01-138)
吴阶平医学基金会临床科研专项资助基金(320.6750.07132)
关键词
胰腺癌
雷帕霉素
MTOR
细胞凋亡
Pancreatic cancer
Rapamycin
Mammalian target of rapamycin
Apoptosis
