Effect of Aldo-keto reductase family 1 member B10 gene silencing on the proliferation and apoptosis of hepatocellular carcinoma cells

Objective To investigate the biological functions and related pathological mechanisms of Aldo-keto reductase family 1 member B10(AKR1 B10) in hepatocellular carcinoma(HCC).Methods The COLE database was used to analyze the mRNA expression of AKR1 B10 in normal liver cells and HCC cell lines.The UALCAN and Oncomine databases were used to analyze the mRNA expression of AKR1 B10 in HCC tissue and adjacent tissue.Western blot was used to investigate the difference in the protein expression of AKR1 B10 between HCC tissue and adjacent tissue.AKR1 B10 in HepG2 and Huh7 cells was knocked down by lentivirus,and the cells were divided into control group and AKR1 B10 knockdown group(which was further divided into shAKR1 B10 1# group and shAKR1 B10 2# group).AnnexinVAPC/PI double staining,colony formation assay,and the method of subcutaneous xenograft tumor were used to observe the apoptosis of HCC cells,colony formation,and the change in the volume of subcutaneous xenograft tumor after AKR1 B10 knockdown.The peroxide-sensitive fluorescent probe DCFH-DA and Western blot were used to measure the changes in ROS and related proteins,such as pATMser1981,γ-H2 AX,c-Caspase-3,and c-RARP in HCC cells.A one-way analysis of variance was used for comparison of continuous data between multiple groups,and the SNK-q test was used for further comparison between two groups.Results The CCLE database showed that the mRNA expression of AKR1 B10 in HCC cells was significantly higher than that in normal liver cells) the Oncomine database suggested that the mRNA expression of AKR1 B10 in HCC tissue was more than 10 times that in adjacent tissue) the UALCAN database showed that the AKR1 B10 gene was highly expressed in HCC tissue.For the six patients in this study,the expression of AKR1 B10 in HCC tissue was significantly higher than that in adjacent tissue.Compared with the control group,the AKR1 B10 knockdown group had a significant reduction in the colony formation ability of HCC cells(P <0.001) and a significant increase in apoptosis rate(P <0.001).Compared with the control group,the AKR1 B10 knockdown group had significant increases in ROS,the markers for DNA damage p-ATMser1981 and γ-H2 AX,and the indices for cell apoptosis c-Caspase-3 and c-RARP.In the BALB-c/Nude mouse model,compared with the control group,the AKR1 B10 knockdown group had a significant reduction in tumor volume since day 13 after allograft transplantation(P <0.05).Conclusion AKR1 B10 may be an effective therapeutic target for the treatment of HCC.