miR-150-5p抑制肝癌细胞的迁移和侵袭及其机制

Inhibitory effect of miR-150-5p on migration and invasion of hepatocellular carcinoma cells and its mechanism

摘要:

目的：探讨miR-150-5p在肝细胞癌（HCC）细胞迁移与侵袭中的作用及其调控机制。 方法：用荧光定量PCR测定miR-150-5p在正常肝细胞系L02及HCC细胞系HepG2中的表达；将HepG2细胞分成两组，分别转染miR-150-5p（miR-150-5p组）与随机序列（对照组），转染后，分别用细胞划痕实验、Transwell小室基质渗透实验检测细胞的迁移和侵袭能力，用Western blot检测细胞基质金属蛋白酶2（MMP2）和基质金属蛋白酶9（MMP9）的蛋白表达。 结果：miR-150-5p的表达量在HepG2细胞系中明显降低，为L02细胞系的0.26倍（P<0.01）。转染后，miR-150-5p组的miR-150-5p水平明显升高，为对照组的9.53倍（P<0.001）；miR-150-5p组的细胞划痕愈合率明显低于对照组（54.63% vs. 87.51%，P<0.01），细胞侵袭数明显少于对照组 （138个vs. 452个，P<0.01）；MMP2与MMP9蛋白表达量均明显低于对照组（0.78 vs. 1.75；0.82 vs. 1.85，均P<0.05）。 结论：miR-150-5p在HCC细胞中表达降低，升高miR-150-5p的表达可抑制HCC细胞的迁移和侵袭，机制可能与其下调MMP2和MMP9表达有关。

Abstract:

Objective: To investigate the effect of miR-150-5p on migration and invasion in hepatocellular carcinoma (HCC) cells and the mechanism. Methods: The miR-150-5p expression in normal hepatic cells and HCC HepG2 cells was determined by real-time PCR. Then, the HepG2 cells were divided into two groups to transfect with miR-150-5p (miR-150-5p group) or scramble sequences (control group), respectively. After transfection, the cell migration and invasion abilities were measured by wound healing assay and Transwell invasion assay, and the protein expressions of matrix metalloproteinase 2 (MMP2) and 9 (MMP9) were examined by Western blot, respectively. Results: The expression level of miR-150-5p in HepG2 cells was significantly reduced, and was 26% of that in normal hepatic cells (P<0.01). After transfection, the miR-150-5p expression level in miR-150-5p group was significantly increased, which was 9.53 times of that in control group (P<0.001). In miR-150-5p group compared with control group, the healing rate of the scratch wound (54.63% vs. 87.51%, P<0.01) and number of invaded cells (138 vs. 452, P<0.01) were all significantly decreased. The protein expression levels of MMP2 and MMP9 (0.78 vs. 1.75; 0.82 vs. 1.85) were all significantly decreased (both P<0.05). Conclusion: The miR-150-5p expression is decreased in HCC cells, and up-regulation of miR-150-5p expression can inhibit the migration and invasion of HCC cells and the mechanism may be associated with its suppressing MMP2 and MMP9 expressions.