miR-580-3p靶向COL11A1调控肝癌细胞凋亡

miR-580-3p targets COL11A1 to regulate apoptosis in hepatocellular carcinoma cells

目的探究miR-580-3p靶向COL11A1调控肝癌细胞凋亡的机制。方法纳入肝细胞癌(HCC)患者42例,实时荧光定量PCR检测肝癌组织、癌旁组织、人正常肝细胞(LO2、QSG-7701)和人肝癌细胞系(HepG2、Huh-7、Li-7、SK-Hep-1)中COL11A1 mRNA和miR-580-3p表达水平。过表达和敲低miR-580-3p后,CCK8法检测肝癌细胞增殖水平,流式细胞术检测细胞的凋亡水平和细胞周期。免疫印迹分析肝癌关键通路Akt/PI3K在miR-580-3p中的作用。结果与癌旁组织比较,肝癌组织COL11A1 mRNA表达升高,miR-580-3p表达降低;与人正常肝细胞系比较,人肝癌细胞系miR-580-3p表达降低、COL11A1 mRNA表达升高(P<0.05)。COL11A1mRNA与miR-580-3p表达呈负相关(P<0.05)。miR-580-3p过表达后,人肝癌细胞的增殖水平、COL11A1 mRNA降低,凋亡率升高;miR-580-3p敲低后上述趋势反之,细胞周期停滞在G1/S期。人肝癌细胞凋亡率COL11A1过表达后降低,敲低后升高。p-Akt、p-PI3K表达在敲低COL11A1或过表达miR-580-3p后降低,过表达COL11A1或敲低miR-580-3p后升高。结论肝癌细胞和肝癌组织miR-580-3p表达水平降低,miR-580-3p/COL11A1轴可能通过调控PI3K/Akt通路影响肝癌细胞凋亡。

Aim To investigate the mechanism of miR-580-3p targeting COL11A1 to regulate apoptosis in hepatocellular carcinoma cells.Methods Forty-two HCC patients were included,The expression levels of COL11A1 mRNA and miR-5803-3p in hepatocellular carcinoma(HCC)tissues,adjacent tissues,human normal hepatocytes(LO2,QSG-7701)and human hepatocellular carcinoma cell lines(HepG2,Huh-7,Li-7,SK-Hep-1)were detected by real-time PCR.After overexpression and knockdown of miR-580-3p.After overexpression and knockdown of miR-580-3p,the proliferation level of hepatocellular carcinoma cells was detected by CCK8 assay,and the apoptosis level and cell cycle were detected by flow cytometry.The role of Akt/PI3K in miR-580-3p was analyzed by Western blotting.Results COL11A1 mRNA expression was elevated and miR-580-3p expression level was decreased in HCC tissues compared with paraneoplastic tissues;miR-580-3p expression was decreased and COL11A1 mRNA expression was increased in human hepatocellular carcinoma cell lines compared with human normal hepatocellular lines(P<0.05).COL11A1 mRNA and miR-580-3p expression were negatively correlated(P<0.05).After miR-580-3p overexpression,the proliferation level,COL11A1 mRNA decreased and apoptosis rate increased in human hepatocellular carcinoma cells;after miR-580-3p knockdown,the above trend was reversed and the cell cycle stalled in G1/S phase.Apoptosis rate of human hepatocellular carcinoma cells decreased after COL11A1 overexpression and increased after knockdown.p-Akt and p-PI3K expression decreased after knockdown of COL11A1 or overexpression of miR-580-3p and increased after overexpression of COL11A1 or knockdown of miR-580-3p.Conclusion The miR-580-3p expression level was reduced in hepatocellular carcinoma cells and hepatocellular carcinoma tissues,and the miR-580-3p/COL11A1 axis may affect apoptosis in hepatocellular carcinoma cells by regulating the PI3K/Akt pathway.