Research On The Expression And Role Of NCEH1 Gene In Hepatocellular Carcinoma Cell Line

BackgroundHepatocellular Carcinoma(HCC)is one of the common malignant tumors of the digestive system,seriously threatening the lives and health of our people.Its incidence ranks sixth in the world as malignant tumors and the fourth in cancer-related deaths.With the continuous improvement of liver cancer surgical techniques,the rate of surgical resection of liver cancer has increased significantly compared with before,and liver transplantation and targeted drugs are also increasingly used in the treatment of liver cancer,but the 5-year overall survival rate of liver cancer is now still no significant improvement.The main reason is that most of the liver cancer patients found in the clinic are in the middle and advanced stages,and it is difficult to undergo radical surgical resection.Therefore,exploring and developing new therapeutic targets is essential to improve the survival time of liver cancer patients.The NCEH1 gene is a gene encoding serine hydrolase,which is closely related to lipoprotein metabolism and the synthesis and secretion of aldosterone.Its activity has been confirmed to be positively correlated with tumor cell invasiveness and is overexpressed in gastric cancer,pancreatic cancer,prostate cancer and other malignant tumors.It is a potential tumor treatment target.However,as of now,there has not been any research report on the expression and role of NCEH1 gene in hepatocellular carcinoma.For this reason,we tried to clarify the role of NCEH1 gene in the occurrence and development of hepatocellular carcinoma in order to discover new therapeutic targets for hepatocellular carcinoma.ObjectiveTo study the expression of NCEH1 gene in hepatocellular carcinoma cells and its effect on the proliferation and apoptosis of hepatocellular carcinoma cells,we hope to clarify the role of NCEH1 gene in hepatocellular carcinoma and its specific molecular mechanism,and provide a potential therapeutic target and theoretical basis for the precise treatment of hepatocellular carcinoma.Research methodsReal-time fluorescence quantitative PCR was used to determine the expression level of NCEH1 gene in hepatocellular carcinoma cell lines and normal liver cells,and then lentivirus-mediated si RNA technology was used to construct hepatocellular carcinoma cell lines with silencing NCEH1 gene.Finally,MTT growth curve method,plate cloning experiment,cell flow cytometry and Annexin V-APC staining method were used to detect the effect of NCEH1 gene on the proliferation,growth,apoptosis and cell cycle of hepatocellular carcinoma cells.ResultThe expression level of NCEH1 gene in the 4 hepatocellular carcinoma cell lines was higher than that of normal liver cell lines(P < 0.05),and the expression was highest in SMMC-7721 cell line.Lentivirus-mediated si RNA technology silenced NCEH1 gene expression in hepatocellular carcinoma SMMC-7721 cell line with an efficiency of 74%.After NCEH1 gene silencing,the results of MTT growth curve showed that the growth rate of hepatocellular carcinoma cells in the NCEH1-si RNA group was considerably lower than that in the NC-si RNA group(F=1262.066,P<0.001).There was no significant difference between the number of clones formed by SMMC-7721 cells in the NCEH1-si RNA group and the number of cell clones in the NC-si RNA group(141±10 vs.144±6,P>0.05).Cell cycle detection showed that the proportion of cells in the G1 and S phases of the NCEH1-si RNA group was considerably lower than that in the NC-si RNA group [G1 stage:(21.87±0.71)% vs.(30±0.93)%,P<0.001;S stage:(33.40±1.22)% vs.(44.50±1.23)%,P<0.001)],while the proportion of cells in the G2/M phase was considerably higher than that of the NC-si RNA group [(44.74±1.17)% vs.(22.94±0.30)%,P<0.001].The apoptotic rate of hepatocellular carcinoma cells in the NCEH1-si RNA group was(6.65 ±0.20)%,while that in the NC-si RNA group was(3.15±0.09)%,that is,the apoptosis rate of NCEH1-si RNA group was 2.11 times that of NC-si RNA group(P<0.001).ConclusionThe expression of NCEH1 gene is elevated in hepatocellular carcinoma cell lines,and silencing its expression can significantly inhibit the growth of hepatocellular carcinoma SMMC-7721 cells and induce G2/M phase arrest and apoptosis,suggesting that NCEH1 gene may be a potential target for hepatocellular carcinoma treatment.