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

摘要:

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。

Abstract:

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 trans－fection 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 as－say results showed that the invasion and migration ability decreased greatly after the transfection with miR-18a ASO（P=0.000），fur－thermore，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 inva－sion and migration of breast cancer cells. miR-18a may become a new target for the regulation of invasion and migration in breast can－cer.