辛伐他汀抑制人乳腺癌MCF-7细胞内多能干细胞标志物Oct3/4 Nanog和Sox-2表达  

Simvastatin inhibits expression of pluripotent markers Oct3/4, Nanog, and Sox-2 in human breast cancer MCF-7 Cells

作　　者：凌桂琴[1] 陈冬波[1] 王保庆[1] 

机构地区：[1]江苏省徐州医学院第二附属医院肿瘤内科,江苏省徐州市221006

出　　处：《中国肿瘤临床》2013年第24期1523-1527,共5页Chinese Journal of Clinical Oncology

基　　金：课题受徐州市科技计划项目(编号:XZZD1255)资助~~

摘　　要：目的:研究辛伐他汀(simvastatin,SIM)对人乳腺癌MCF-7细胞内多能干细胞标志物Oct3/4、Nanog和Sox-2表达的影响。方法:应用实时荧光定量聚合酶链式反应(qRT-PCR)技术、免疫荧光染色方法,流式细胞仪和免疫印迹(Western blot)方法检测SIM对MCF-7细胞内多能干细胞标志物表达的影响。结果:qRT-PCR结果显示,10、50和100μmol/L SIM作用于MCF-7细胞48h后,能显著抑制细胞内Oct3/4、Nanog和Sox-2基因表达,与对照组相比,差异有统计学意义(P<0.05),而SIM 1μmol/L浓度组和对照组相比差异无统计学意义(P>0.05)。SIM 50、100μmol/L浓度组和10μmol/L浓度组相比,抑制Oct3/4和Nanog的表达差异有统计学意义(P<0.05),而对Sox-2的表达抑制,SIM 10、50μmol/L和100μmol/L各浓度组间差异无统计学意义(P>0.05)。免疫荧光染色显示经10μmol/L SIM处理48 h后MCF-7细胞核内Oct3/4、Nanog和Sox-2蛋白表达减弱,部分细胞核无表达。流式细胞检测显示MCF-7细胞经10μmol/L SIM处理48 h后,Oct3/4阳性细胞数、Nanog阳性细胞数和Sox-2阳性细胞数显著减少(P<0.05),Western blot进一步证实经10μmol/L SIM处理48 h后MCF-7细胞核内Oct3/4、Nanog和Sox-2蛋白表达显著减少(P<0.05)。结论:SIM在体外能有效地抑制人乳腺癌MCF-7细胞内多能干细胞标志物Oct3/4、Nanog和Sox-2的表达,为SIM应用于癌症治疗提供实验依据。Objective：To investigate the effect of simvastatin on expression of pluripotent markers Oct3/4, Nanog, and Sox-2 in human breast cancer MCF-7 cells. Methods：Quantitative reverse transcription-polymerase chain reaction （qRT-PCR）, immunofluo-rescent staining, flow cytometry, and Western blot were used to detect the expression of pluripotency markers Oct3/4, Nanog, and Sox-2 in human breast cancer MCF-7 cells treated with different doses of simvastatin. Results：qRT-PCR revealed the more signifi-cant inhibition of gene expressions of Oct3/4, Nanog, and Sox-2 in human breast cancer MCF-7 cells when subjected to high doses of simvastatin （10, 50, and 100 μmol/L） compared with the control group （P〈0.05）. By contrast, no significant difference was observed be-tween the expressions of low-dose simvastatin treatment （1 μmol/L） and control （P〉0.05）. The inhibitory effect of simvastatin on the gene expressions of Oct3/4 and Nanog was more significantly apparent at 50 and 100 μmol/L dosages than at 10 μmol/L （P〈0.05）. By contrast, no significant difference in the inhibitory expression of Sox-2 was observed among the three high-dose treatments （P〉0.05）. Between the two higher-dose treatments （50 and 100 μmol/L）, no significant difference in the inhibitory expressions of Oct3/4, Nanog, and Sox-2 in MCF-7 cells was found. Meanwhile, in the 10 μmol/L simvastatin treatment, immunoflurescent staining showed a marked reduction in the protein expression of all three pluripotent markers in MCF-7 cells, and flow cytometry demonstrated a decrease of Oct3/4-, Nanog-, and Sox-2-positive cells （P〈0.05）. Western blot further revealed that the protein expression of Oct3/4, Nanog, and Sox-2 in MCF-7 cells was significantly declined by the same simvastatin dose （P〈0.05）. Conclusion： Simvastatin can inhibit the expression of pluripotent markers Oct3/4, Nanog, and Sox-2 in human breast cancer MCF-7 cells, proving the anti-cancer properties of simvastatin.

关 键 词：辛伐他汀 乳腺癌MCF-7细胞 多能干细胞标志物 OCT3 4 NANOG Sox-2 

分 类 号：R737.9[医药卫生—肿瘤]