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

摘要:

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-93 ASO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。

Abstract:

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 overex－pressed in the human breast cancer. Reducing expression of miR-93 can effectively inhibit the growth of breast cancer cells and pro－mote apoptosis . miR-93 may become a new target for the regulation of gene expression in breast cancer.