miR-1470对人肝癌细胞增殖和凋亡的影响

目的 探讨miR-1470对人肝细胞癌增殖和凋亡的影响。 方法 采用实时荧光定量PCR（quantitative real-time PCR，qPCR）检测人正常肝细胞株（LO2）和人肝癌细胞株（HepG2、Hep3B、Huh7）中miR-1470的差异表达；采用慢病毒转染肝癌细胞株，过表达（HepG2细胞株，过表达组）或敲减miR-1470（Huh7细胞株，抑制组）后，qPCR检测各组细胞miR-1470的表达；CCK8法检测细胞增殖改变；流式细胞术检测细胞凋亡情况。结果 miR-1470在肝癌细胞系HepG2、Hep3B和Huh7中的表达水平高于正常肝细 胞系LO2，HepG2、Hep3B、Huh7三种细胞系相对正常细胞表达量分别为：2.304±0.366、2.851±0.508、 5.768±0.445（均P＜0.05）。CCK8实验证实，过表达miR-1470组OD值在72 h显著高于对照组（P＜0.05），而抑制组显著低于对照组（P＜0.05）。流式细胞分析结果显示过表达miR-1470抑制肝癌细胞的凋亡（P＜0.05）；而miR-1470抑制组对比对照组，凋亡细胞显著增多（P＜0.05）。结论 miR-1470促进肝癌细胞增殖，抑制肝癌细胞凋亡。

Effects of miR-1470 on cell proliferation and apoptosis in human hepatocellular carcinoma cells #br#

Objective To investigate the effects of miR-1470 on the proliferation and apoptosis of hepatocellular carcinoma (HCC) cell. Methods The differential expressions of miR-1470 in normal liver cell line (LO2) and HCC cell lines (HepG2, Hep3B, Huh7) were detected by quantitative real-time PCR (qPCR). Lentivirus transfection was used to up-regulate (HepG2 cell line) or down-regulate miR-1470 (Huh7 cell line) in HCC cell lines and the expression levels of miR-1470 in transfected cell lines were detected by qPCR. CCK8 assay was used to detect cell proliferation, and flow cytometry was used to detect cell apoptosis. T-test was used for comparison between two groups, and one-way ANOVA was used for comparison among multiple groups. Results The expression levels of miR-1470 in HCC cell lines HepG2, Hep3B and Huh7 were higher than that in normal liver cell line LO2. Compared with LO2, the relative expression levels of the three HCC cell lines HepG2, Hep3B and Huh7 were 2.304±0.366, 2.851±0.508, 5.768±0.445 respectively (all P<0.05). CCK8 experiment confirmed that OD values were significantly higher in the overexpressed miR-1470 groups than that in the control group at 72 h (P<0.05), while which were significantly lower in the inhibitory groups than that in the control group (P<0.05). Flow cytometry showed that overexpression of miR-1470 inhibited the apoptosis of HCC cells (P<0.05). Compared with the control group, the apoptotic cells increased significantly in miR-1470 down-regulated groups (P<0.05). Conclusion miR-1470 promotes cell proliferation and inhibits cell apoptosis in HCC.