Department of Hepatobiliary Surgery, Guangdong General Hospital, Guangdong Academy of Medical Sciences, Guangzhou, Guangdong, China.
IUBMB Life. 2018 Jun;70(6):468-478. doi: 10.1002/iub.1715. Epub 2018 Apr 20.
The miR-490-3p was transfected into HepG2 cells to explore the correlation between miR-490-3p and hepatocellular carcinoma cell proliferation, apoptosis, and autophagy and its downstream target gene ATG7. Then we could possibly provide a mechanism for the treatment of hepatocellular carcinoma. MiR-490-3p was screened out by fold change > 4 and P < 0.01 using gene microarray data. The expression level of miR-490-3p was tested by qRT-PCR and the prognosis analysis was achieved by using TCGA data. The cell proliferation was tested via colony formation assay and CCK-8 after the miR-490-3p mimics were transfected into HepG2 cells; the variations of cell cycle and apoptosis was examined by flow cytometry assay; the number of autophagosome was observed by electron microscopy and the changes of autophagy-relative protein LC-II and LC-I as well as their ratio was tested by western blot. MiR-490-3p is low expressed in hepatocellular carcinoma cell lines and tissues. The results of TCGA showed that miR-490-3p high expression indicated better prognosis. After HepG2 cells were transfected with miR-490-3p mimics, cell viability was increased, cell proliferation was enhanced, cell cycle was blocked in G0/G1 phase, cell apoptosis rate was increased, the number of autophagosomes was reduced, autophagy-associated protein LC-II was decreased, and LC-I was increased and their ratio was decreased. After 3-MA was added, cell proliferation was declined, cell apoptosis rate was increased. Besides, the autophagy was inhibited by knocking down the ATG7, which promoted the cell apoptosis. MiR-490-3p could suppress cell proliferation, retard cell cycle and upgrade cell apoptosis by inhibiting autophagy in HCC cells via targeting ATG7. © 2018 IUBMB Life, 70(6):468-478, 2018.
miR-490-3p 转染 HepG2 细胞，探讨 miR-490-3p 与肝癌细胞增殖、凋亡和自噬及其下游靶基因 ATG7 的相关性。然后，我们可能为肝癌的治疗提供一种机制。通过基因微阵列数据筛选出差异倍数>4 和 P<0.01 的 miR-490-3p。采用 qRT-PCR 检测 miR-490-3p 的表达水平，利用 TCGA 数据进行预后分析。miR-490-3p 模拟物转染 HepG2 细胞后，通过集落形成实验和 CCK-8 检测细胞增殖；通过流式细胞术检测细胞周期和凋亡的变化；通过电子显微镜观察自噬体的数量，通过 Western blot 检测自噬相关蛋白 LC-II 和 LC-I 的变化及其比值。miR-490-3p 在肝癌细胞系和组织中低表达。TCGA 的结果表明，miR-490-3p 高表达预示着更好的预后。转染 miR-490-3p 模拟物后，HepG2 细胞活力增加，细胞增殖增强，细胞周期阻滞在 G0/G1 期，细胞凋亡率增加，自噬体数量减少，自噬相关蛋白 LC-II 减少，LC-I 增加，其比值降低。加入 3-MA 后，细胞增殖下降，细胞凋亡率增加。此外，通过敲低 ATG7 抑制自噬，促进细胞凋亡。miR-490-3p 通过靶向 ATG7 抑制自噬，可抑制 HCC 细胞增殖，阻滞细胞周期，促进细胞凋亡。
