辛伐他汀和NONO对乳腺癌细胞增殖的影响

The effects of simvastatin and NONO on the proliferation of breast cancer cells

目的:探讨辛伐他汀和NONO(non-POU-dormain-containing,octamerbinding protein)对乳腺癌MCF-7细胞增殖的影响及其可能的作用机制。方法 :将靶向NONO基因的小干扰RNA(small interfering RNA,si RNA)片段si RNA-NONO和NONO过表达重组载体pc DNA4/TO-NONO分别转染至乳腺癌MCF-7细胞后,分别应用实时荧光定量PCR、蛋白质印迹法和细胞计数法检测NONO m RNA和蛋白的表达及细胞的增殖情况。辛伐他汀(20μmol/L)处理MCF-7细胞后,分别应用细胞计数法、实时荧光定量PCR和蛋白质印迹法分别检测细胞的增殖、NONO m RNA及蛋白的表达水平。辛伐他汀处理pc DNA4/TO-NONO转染组MCF-7细胞后,应用细胞计数法检测细胞的增殖情况。实时荧光定量PCR和组织总胆固醇酶法检测si RNA-NONO转染组MCF-7细胞中3-羟基-3-甲基戊二酰辅酶A还原酶(3-hydroxy-3-methylglutaryl-Co A reductase,HMGCR)m RNA和胆固醇水平。结果 :si RNA-NONO转染组MCF-7细胞中NONO m RNA和蛋白的表达水平低于阴性对照组[MCF-7细胞转染si RNA negative contro(lsi RNANC)](P<0.01,P<0.05),pc DNA4/TO-NONO转染组MCF-7细胞中NONO m RNA和蛋白的表达水平高于转染空载体pc DNA4/TO的对照组(P值均<0.01)。si RNA-NONO转染组MCF-7细胞的增殖能力明显低于si RNA-NC转染组(P<0.05),而pc DNA4/TO-NONO转染组MCF-7细胞的增殖能力明显高于转染空载体pc DNA4/TO的对照组(P<0.05)。辛伐他汀(20μmol/L)处理组MCF-7细胞的增殖能力和NONO mR NA及蛋白的表达水平均明显低于辛伐他汀未处理的对照组(P<0.05,P<0.01,P<0.05)。pc DNA4/TO-NONO转染组细胞经辛伐他汀处理后,细胞的增殖能力明显低于未用辛伐他汀处理的细胞(P<0.01)。si RNANONO转染组MCF-7细胞中HMGCR m RNA和胆固醇水平均明显低于si RNA-NC转染组(P值均<0.01)。结论 :辛伐他汀可以抑制MCF-7细胞的增殖,可能与抑制NONO的表达有关。

Objective：To investigate the effects of simvastatin and non-POU-dormain-containing,octamer-binding protein（NONO） on the proliferation of breast cancer MCF-7 cells,and to explore its possible mechanism.Methods：The expression levels of NONO mRNA and protein and the proliferation of breast cancer MCF-7 cells after transfection with NONO gene-targeted small interfering RNA（siRNA）（siRNA-NONO） and the NONO over-expression recombination vector pcDNA4/T0-NONO were detected by real-time fluorescence quantitative PCR,Western blotting,and cell counting,respectively.The proliferation and expression levels of NONO mRNA and protein of breast cancer MCF-7 cells after treatment with simvastatin（20 μmol/L） were detected by cell counting,real-time fluorescence quantitative PCR,and Western blotting,respectively.The proliferation of MCF-7 cells after treatment with simvastatin in pcDNA4/TO-NONO transfection group was determined by cell counting.The expression level of 3-hydroxy-3-methylglutaryl-CoA reductase（HMCCR） mRNA and the concentration of cholesterol were examined by real-time fluorescence quantitative PCR and tissue total cholesterol assay,respectively.Results：The expression levels of NONO mRNA and protein of MCF-7 cells in siRNA-NONO transfection group were lower than those in siRNA negative control（siRNA-NC） transfection group（P〈0.01,P〈0.05）.The expression levels of NONO mRNA and protein of MCF-7cells in pcDNA4/TO-NONO transfection group were higher than those in the empty vector pcDNA4/TO transfection group（both P〈0.01）.The ability of proliferation of MCF-7 cells in siRNA-NONO transfection group was lower than that in siRNA-NC group（P〈0.05）,while the ability of proliferation of MCF-7 cells in pcDNA4/TO-NONO transfection group was higher than that in the empty vector pcDNA4/TO transfection group（P〈0.05）.The ability of proliferation and the expression levels of NONO mRNA and protein of MCF-7 cells in simvastatin（20 μmol/L） treatment group were lower than those in the control group（MCF-7cells without simvastatin treatment）（P〈0.05,P〈0.01,P〈0.05）.The ability of proliferation of MCF-7 cells after treatment with simvastatin in pcDNA4/TO-NONO transfection group was lower than that in the group without simvastatin treatment（P〈0.01）.The expression levels of HMCCR mRNA and cholesterol of MCF-7 cells in siRNA-NONO transfection group were lower than those in the siRNA-NC transfection group（all P〈0.01）.Conclusion：Simvastatin can suppress the proliferation of breast cancer MCF-7 cells,and this effect may be related to the inhibition of NONO expression.