反义miR-18a对乳腺癌细胞侵袭迁移的影响

Effect of antisense miR-18a on invasion and migration in breast cancer cells

目的:分析miR-18a在乳腺癌组织中的表达情况;体外研究miR-18a反义寡核酸(antisense oligonucleotide,ASO)对乳腺癌细胞侵袭和迁移的影响。方法:运用荧光定量PCR检测108例乳腺癌组织及对应癌旁乳腺组织中miR-18a的表达;通过miR-18a ASO降低乳腺癌细胞中miR-18a的表达,采用Transwell实验和划痕实验观察miR-18a ASO对乳腺癌细胞侵袭和迁移能力的影响,同时运用Western blot的方法检测侵袭相关蛋白的表达变化。结果:在108例乳腺癌病例中,50.93%(55/108)的乳腺癌组织miR-18a表达明显高于对应正常组织(t=28.68,P=0.000);与空白对照组和转染随机ASO组相比,miR-18a ASO可以明显降低miR-18a的表达(F=321.67,P=0.001;F=563.28,P=0.000);Transwell实验和划痕实验结果显示转染miR-18a ASO后,乳腺癌细胞的侵袭迁移能力明显下降(P=0.000),同时相关侵袭蛋白基质金属蛋白酶(matrix metalloproteinase,MMP)2和MMP9表达下降(P=0.000)。结论:miR-18a在乳腺癌组织中表达上调,降低miR-18a的表达,可有效抑制乳腺癌细胞的侵袭和迁移。miR-18a有可能成为乳腺癌侵袭转移调控的新靶点。

Objective：To investigate the expression of miR-18a in breast cancer tissues and to explore the regulative effects of miR- 18a antisense oligonucleotide（ASO） on the invasion and migration of breast cancer cells in vitro. Methods：The expression of miR- 18a in 108 breast cancer tissues and their adjacent breast tissues were detected by real-time quantitative PCR method. After transfection with miR-18aASO,the biological effects of miR-18a on breast cancer cells was measured by Transwell assay and wound healing assay and invasion-related protein expression was analyzed by Western blot. Results：miR-18a was found to be overexpressed in 50.93%（55/108） of the breast cancer cases（t=28.68,P=-0.000）, miR-18a expression in breast cancer cells（transfection with miR-18a ASO） was significantly less than that of control group（F=321.67,P=0.001 ;F=563.28,P=-0.000）. Transwell and wound healing assay results showed that the invasion and migration ability decreased greatly after the transfection with miR-18a ASO （P=0.000）, fursay results showed that the invasion and migration ability decreased greatly after the transfection with miR-18a ASO （P=0.000）, furthermore,down-regulation of miR-18a resulted in obvious inactivation of matrix metalloproteinase （MMP）2 and MMP9 （P=0.000）. Conclusions：miR-18a is overexpressed in human breast cancer. Reducing the expression of miR-18a can effectively inhibit the invasion and migration of breast cancer cells, miR-18a may become a new target for the regulation of invasion and migration in breast cancer.