miRNA-132通过Hedgehog信号通路对肝癌细胞凋亡的作用机制*

目的：探究miRNA-132（miR-132）通过Hedgehog 信号通路对肝细胞癌细胞凋亡的机制。方法：用miR- 132 类似物转染人肝癌HepG2 细胞，将样本分为空白组（ 无转染HepG2）、NC组（HepG2 转染miR-132-NC）、 YJ 组（HepG2 转染miR-132 mimic）。通过qRT-PCR 检测miR-132 在HepG2 细胞中的表达量，采用CCK-8 法、流 式细胞仪、Transwell 小室、免疫印迹检测细胞增殖、凋亡、侵袭能力以及Shh 蛋白表达。结果：PCR检测结果显示， 3 组对比，YJ 组HepG2 细胞中miR-132 表达量最高，说明转染成功；CCK-8 检测结果显示，YJ 组HepG2 细胞增 殖数量最低，空白组HepG2 细胞的增殖与NC组相似，皆高于YJ 组；流式细胞检测结果显示，与NC组及空白组 比较，YJ 组HepG2 细胞凋亡数量最多 ；Transwell 小室检测结果显示， YJ 组细胞侵袭数量与空白组和NC组相比 明显降低，空白组细胞侵袭数量与NC组相比数据接近；免疫印迹检测结果显示，YJ 组Shh 蛋白相对表达量与空 白组和NC组比明显降低。结论：过表达miR-132 降低Hedgehog 信号通路，可促进人肝癌HepG2 细胞凋亡作用。

Mechanism of miRNA-132 on apoptosis of hepatoma cells through Hedgehog signaling pathway

Objective To explore the mechanism of miRNA-132（miR-132） on apoptosis in hepatoma cells through Hedgehog signaling pathway. Methods Human liver cancer HepG2 cells were transfected with miR-132 mimics， and the samples were divided into blank group， NC group transfected with miR-132-NC， and YJ group transfected with miR-132 mimics. QRT-PCR was used to detect the expression of miR-132 in human hepatocellular carcinoma HepG2 cells. QRT-PCR， CCK-8 method， flow cytometry， Transwell cell， and Western blotting were used to detect cell proliferation， apoptosis， invasive ability and Shh protein expression. Results PCR test results showed that the expression of miR-132 was the highest in HepG2 cells in the three groups compared with YJ group， indicating successful transfection ； CCK-8 test results showed that the proliferation of HepG2 cells in the YJ group was the lowest in the three groups. The proliferation of HepG2 cells in the blank group was similar to that in the NC group， which was higher than that in the YJ group. The results of flow cytometry showed that compared with the NC group and the blank group， the number of apoptosis of HepG2 cells in the YJ group was the most， the gap between the number of cells in the blank group and the NC group was smaller. The results of the transwell cell test showed that the number of cell invasions in the YJ group was significantly less than that in the blank group and the NC group， the number of cell invasion in the blank group was close to that in the NC group. Western blotting results showed that the relative expression level of Shh protein in YJ group was significantly higher than that in blank group and NC group， and the relative expression level of Shh protein in blank group was close to that in NC group. Conclusion Overexpression of miR-132 reduces Hedgehog signaling pathway and promotes apoptosis of human liver cancer HepG2 cells.