反义miR-30d对乳腺癌细胞侵袭迁移能力的影响

The Effect of Antisense miR-30d on Invasion and Migration in Breast Cancer Cells

目的分析miR-30d在乳腺癌组织中的表达情况,体外研究反义miR-30d寡核苷酸(miR-30d 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正常癌旁组织中miR-30d的表达;通过miR-30d ASO降低乳腺癌细胞中miR-30d的表达,采用Transwell实验和划痕实验观察miR-30d ASO对乳腺癌细胞侵袭和迁移能力的影响,运用Western blot方法检测侵袭相关蛋白基质金属蛋白酶(MMP)-2和MMP-9蛋白的表达变化。结果乳腺癌组织中miR-30d基因表达水平明显高于对应癌旁组织(P<0.05);与空白对照组和无义干扰组相比,miR-30d ASO可以显著降低乳腺癌细胞miR-30d的表达(P<0.05);Transwell实验和划痕实验结果显示转染miR-30d ASO后,乳腺癌细胞的侵袭及迁移能力明显下降,同时MMP-2和MMP-9表达下降(P<0.05)。结论miR-30d在乳腺癌组织中表达上调,降低miR-30d的表达可有效抑制乳腺癌细胞的侵袭和迁移。miR-30d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-30d in breast cancer tissues and demonstrate the regula-tive effects of miR-30d ASO on the invasion and migration of breast cancer cells in vitro. Methods The expression levels of miR-30d in 108 breast cancer tissues and their adjacent tissues were detected by real-time quantitative PCR method. Af-ter transfection with miR-30d ASO, the biological effects of miR-30d on in breast cancer cells was measured by transwell as-say and wound healing assay. The expressions of matrix metalloproteinase （MMP）-2 and MMP-9 were analyzed by Western blot assay. Results The expression level of miR-30d was found to be over-expressed in breast cancer tissues（P＆lt;0.05）. Compared with control group and nonsense interference group, the miR-30d expression was significantly decreased in breast cancer cells（transfection with miR-30d ASO）. Results of transwell and wound healing assay showed that the invasion and mi-gration ability decreased significantly after transfection with miR-30d ASO, and expressions of MMP-2 and MMP-9 were down-regulated （P＆lt;0.05）. Conclusion miR-30d was over-expressed in human breast cancer. The invasion and migration of breast cancer cells can be effectively inhibited by decreasing the expression of miR-30d. miR-30d may become a new tar-get for the regulation of invasion and migration in breast cancer.