The Expression Of Panx1 In Hepatocellular Carcinoma And Its Effect On The Growth Of HCC

Objective Although the clinical diagnosis and surgical treatment of liver cancer have made great progress,the mortality rate of liver cancer still ranks in the top three.In the early stage of liver cancer,it is not easy to be detected.Most patients have undergone intrahepatic metastases or even distant metastases at the time of diagnosis.Therefore,it is important and urgent to develop new diagnostic and prognostic factors for hepatocellular carcinoma.Recent studies have found that Panx1(Pannexin1)is closely related to the development of tumor and malignant metastasis.Panx1 channel inhibitors can significantly reduce the metastasis efficiency of breast cancer.The knockout of low Panx1 gene can obviously inhibit the proliferation of glioma cells.The expression of Panx1 is negatively correlated with the proliferation of gallbladder cancer.In the development of these tumors,Panx1 may play a role in inhibiting cancer or promoting cancer as a oncogene,and the function and molecular mechanism of Panx1 gene in liver cancer has not been elucidated.The purpose of this study is to explore the expression of Panx1 in liver cancer and its effect on the proliferation of liver cancer,to explore the factors related to the growth of liver cancer,to find new targets for prevention and treatment,and to provide possible help for the treatment and prognosis of liver cancer.Methods1.In the TCGA database,we searched for the RNA expression level of Panx1 gene in hepatocellular carcinoma.2.Immunohistochemical staining was used to detect the expression of Panx1 protein in63 cases of hepatocellular carcinoma(cancer and paracancerous).3.Human hepatoma cell lines Hep3 B,huh7,97 L,97H,LM3 were detected by Western blotting technique.In the cell lines 97 L and LM3 with relatively low Panx1 expression,the panx1 overexpression gene was introduced into the 97 L and LM3 cells by the lentivirus technique to construct the Panx1 gene overexpression stable transformant.4.Western blot analysis was used to detect the expression levels of Panx1 and PTEN in hepatoma stable transfected cells 97 L and LM3.5.CCK8 technology was used to detect the proliferation of 97 L and LM3 in stable cell lines.The effect of Panx1 gene overexpression on proliferation of 97 L and LM3 cells was detected.6.LM3 cells were subcutaneously implanted in nude mice to detect the effect of overexpression of Panx1 gene on subcutaneous tumor formation in mice.7.In Panx1 knockout mice and wild type mice,the effect of knocking out Panx1 on the tumorigenesis of hepatoma cells in Hepa1-6 mice was detected.Results1.In the TCGA data,we detected that Panx1 gene was lowly expressed in hepatocellular carcinoma,and Panx1 gene expression was higher in paracancerous tissues.2.In 63 cases of hepatocellular carcinoma and paracancerous tissues,the expression level of Panx1 protein in hepatocellular carcinoma was significantly lower than that in paracancerous tissues.3.In the hepatoma cell line,Panx1 was found to have low expression in 97 L and LM3 cells.Overexpression of Panx1 gene increases PTEN protein expression.4.In vitro cell proliferation test,compared with the control group,overexpression of Panx1 gene significantly inhibited the proliferation of 97 L and LM3 stable cells.5.Compared with the control group,the overexpression of Panx1 gene could significantly inhibit the ability of LM3 cells to be tumorigenic in nude mice,and the subcutaneous tumor volume decreased significantly in the nude mice.6.Compared with the wild-type mice,Panx1 knockout mice may significantly promote the proliferation of Hepa1-6 cells in mice.The subcutaneous tumor volume was significantly greater than that of the wild-type group.Conclusions1.We detected that the expression of Panx1 in liver cancer tissues was significantly lower than that of normal tissues beside liver cancer.The expression of L02 in normal liver cells of Panx1 was significantly higher than that of other hepatoma cells,and the expression of panx1 was significantly different.2.Overexpression of Panx1 can significantly inhibit the proliferation of hepatoma cells in vitro and in vivo.Its expression may play a role in inhibiting the growth of liver cancer,which may be involved in the occurrence,development and prognosis of liver cancer.