shRNA沉默ADAM17基因对人乳腺癌细胞MCF-7增殖的影响

Effects of shRNA-targeted ADAM17 gene on proliferation of human breast cancer cells MCF-7

关键词:  解聚素-金属蛋白酶17  短发夹RNA  乳腺癌  增殖

Key words:  ADAM17  shRNA  breast cancer  proliferation

摘要:

目的　利用shRNA干扰沉默人乳腺癌细胞MCF-7的解聚素-金属蛋白酶17（adisintegrin and metalloproteinases 17，ADAM17）基因，观察其对细胞增殖的影响。方法　针对ADAM17基因设计合成具有特异性的ADAM17-shRNA，经脂质体LipofectamineTM 2000转染MCF-7细胞。实验设对照组（空白PBS）、干扰组（转染干扰无义序列ADAM17-shNC）、实验组（转染ADAM17-shRNA），采用Realtime PCR检测各组细胞ADAM17 mRNA的表达水平，四甲基偶氮唑盐比色法（MTT）检测细胞的增殖活性，流式细胞仪检测细胞周期变化。结果　实验组ADAM17 mRNA的相对表达量显著低于对照组和干扰组，差异有统计学意义（P<0.05）；实验组的MCF-7细胞增殖活性较其他两组明显降低，差异有统计学意义（P<0.05）；实验组的MCF-7细胞进入S期（23.60±1.09）%和G₂/M期（6.30±0.82）%的比例降低，绝大多数细胞停留在G₀/G₁期（65.17±1.35）%，细胞周期延缓，与对照组、干扰组相比较差异具有显著性意义(P<0.05)。结论　ADAM17-shRNA对人乳腺癌细胞MCF-7的ADAM17基因具有沉默作用，从而抑制MCF-7细胞的增殖活性，延缓细胞周期进展。

Abstract:

Objective　 To investigate the effect of ADAM17-shRNA on the proliferation of human breast Cancer MCF-7cells by shRNA silence ADAM17 gene.Methods　The specific ADAM17-shRNA,which was designed for ADAM17gene,They were transfected into MCF-7 cells by lipofectamine TM2000. Experiment groups were divided into control group(PBS), Interference group(was transfected with ADAM17-shNC) and experience group(was transfected with ADAM17-shRNA). ADAM17 mRNA expression were detected by Real-time PCR, The proliferative ability of MCF-7 cells were detected by MTT, Cell cycle distribution of MCF-7 cells was analyzed by FCM.Results　The relative expression of ADAM17mRNA in experience group was significantly lower than control groups and interferencegroup.So the difference was statistically significant (P<0.05), the proliferation of MCF-7 cells in experiencegroup was significantly reduced than control groups and Interference group. So the difference was statistically significant (P<0.05), with experiencegroup, the proportion of MCF-7 cells into the S phase (23.60±1.09)% and G₂/M phase (6.30±0.82)% was lowered, with the vast majority of cells remaining in the G₀/G₁phase (65.17±1.35)%. Compared with the control group and Interference group, so the difference was statistically significant (P<0.05).Conclusions 　ADAM17-shRNA could effectively silence the expression of ADAM17 gene in human breast cancer MCF-7 cells, so that the proliferation of MCF-7 cells is inhibited and the cell cycle is delayed.