miR-93反义核苷酸对乳腺癌细胞增殖及凋亡的影响

Effects of miR-93 antisense oligonucleotides on cell proliferation and apoptosis in breast cancer

目的:研究miR-93在乳腺癌组织中的表达情况,并分析miR-93反义核苷酸(antisense oligonucleotide,ASO)对乳腺癌细胞增殖和凋亡的影响。方法:运用荧光定量PCR定量检测120例乳腺癌组织及对应癌旁组织中miR-93的表达;通过miR-93ASO降低乳腺癌细胞中miR-93的表达,利用MTT、细胞克隆形成实验、流式细胞仪观察miR-93 ASO对乳腺癌细胞产生的生物学效应。实验分为3组:空白对照组,无义干扰组和miR-93 ASO组。结果:在120例乳腺癌病例中,63.33%(76/120)的乳腺癌组织miR-93表达明显高于对应癌旁组织(P<0.05);miR-93 ASO可以显著降低miR-93的表达(P<0.05);MTT实验结果显示乳腺癌细胞转染miR-93 ASO 24、48 h和96 h后的细胞存活数量均明显低于空白对照组和无义干扰组(P<0.05);克隆形成实验结果显示miR-93 ASO组克隆形成率比空白对照组和无义干扰组显著降低(P<0.05);流式细胞仪检测结果显示转染miR-93ASO组较2个对照组细胞凋亡指数明显增加(P<0.05);另外,降低miR-93的表达后发现Bcl2的mRNA和蛋白均明显下降(P<0.05)。结论:miR-93在乳腺癌组织中表达上调,降低miR-93的表达可有效抑制乳腺癌细胞生长、促进细胞凋亡。miR-93有可能成为乳腺癌基因表达调控的新靶点。

Objective：To investigate the expression of miR-93 in breast cancer tissues and to explore the regulative effects of miR-93 antisense oligonucleotide（ASO） on the proliferation and apoptosis of breast cancer cells. Methods：Expression of miR-93 in 120 breast cancer tissues and their adjacent tissues was detected by real-time quantitative PCR. After transfection with miR-93 ASO, biological effects of miR-93 on breast cancer cells were measured by MTT assay, colony formation experiment and flow cytometry. Results：Expression of miR-93 was higher in 63.33%（76/120） breast cancer tissues than in adjacent tissues（P〈0.05）, miR-93 ASO could reduce the expression of miR-93 significantly（P〈0.05）. MTT assay results showed that breast cancer cells survival rate at 24,48,96 h decreased greatly after transfection with miR-93 ASO（P〈0.05）. Clone formation assay revealed that colony formation rate was significantly lower in miR-93 ASO group than in blank control group and nonsense interference group （P〈0.05）. Flow cytometry indicated that the apoptotic index was significantly higher in miR-93 ASO group than in two control groups（P〈0.05）. In addition, Bcl2 mRNA and protein levels were significantly decreased after reducing expression of miR-93. Conclusions：miR-93 is overexpressed in the human breast cancer. Reducing expression of miR-93 can effectively inhibit the growth of breast cancer cells and promote apoptosis, miR-93 may become a new target for the regulation of gene expression in breast cancer.