Upregulation Of K19 Expression By HBx Protein Promotes Hepatocellular Carcinoma Invasion And Metastasis And Postoperative Recurrence

Hepatocellular carcinoma(HCC)ranks sixth among gastrointestinal tumors worldwide,and is a gastrointestinal tumor associated with cancer death.Hepatitis B virus(HBV)is an important risk factor for hepatocellular carcinoma.Hepatitis B virus X protein(HBx)is a multifunctional protein encoded by the HBV virus gene,which plays an important role in virus replication,gene transcription and transduction of key signal pathways,and can regulate tumor cells A variety of biological functions such as proliferation,chemotherapy resistance,and invasion and metastasis are closely related to the recurrence and metastasis of liver cancer.HCC cells positively express K19(Keratin19,K19)with the characteristics of cancer stem cells.According to the theory of liver cancer stem cells,hepatic progenitor cells(HPC)play the role of stem cells,and cytokeratin 19 is a cancer stem cell-specific molecule Markers,in terms of HCC biology,such as recurrence and metastasis,the positive expression of HCC is more likely than the negative expression of HCC,which has become a major difficulty in the prevention and treatment of liver cancer.In order to study the role of HBx protein in regulating K19 expression in HCC recurrence and metastasis,HCC cell lines which were trasfected with the HBx were uesed to regulate HBx expression,and performed functional experiments on the transfected cells to observe the effect of HBx on HCC cells.Using SMMC-7721 and HepG2 hepatocellular carcinoma cell lines stably transfected with HBx to regulate the K19 gene,functional experiments were conducted to further explore the role of HBx in regulating K19 to promote the invasion and metastasis of liver cancer cells.In addition,we collected hepatocellular carcinoma tissue specimens,and detected the correlation between the expression of HBx and K19 in liver cancer tissues by RT-PCR,Western-blot,and immunohistochemistry to verify the relationship between the two;analyze the expression levels of HBx and K19 The relationship between the level and age of patients with hepatocellular carcinoma,age,sex,liver cirrhosis,and degree of liver cancer differentiation,preoperative AFP levels,vascular invasion,HBV infection,and TNM staging.To evaluate the clinical significance of HBx and K19 in liver cancer recurrence,metastasis and prognosis.Objective(s):1.The effects of HBx and K19 on HCC invasion and migration were clarified.2.To investigate the expression of HBx and K19 in HCC,and to analyze its relationship with clinicopathological factors and prognosis of patients with hepatocellular carcinoma.Methods:1.SMMC-7721 and HepG2 were transfected into LV K19,K19 shRNA and LV HBx to regulate related genes.2.After successfully transfecting SMMC-7721 and HepG2 cells with HBx and K19,through the invasion and scratch experiments,it is clear whether HBx and K19 promote the migration and invasion ability of HCC cells.3.The liver cancer tissue specimens were analyzed by immunohistochemistry for the correlation between the expression of HBx and K19 in tumor tissues and surrounding tissues of patients,and the relationship with the clinicopathological factors,recurrence and metastasis and survival time of patients with liver cancer.Results:1.In HCC,HBx can promote tumor cell invasion and migration through K19.2.In immunohistochemistry experiments,the expression of HBx and K19 in HCC has a correlation,which is closely related to the degree of HCC differentiation,HBV positive,higher AFP levels,vascular invasion and TNM staging factors.Conclusion(s):1.HBx can promote HCC invasion and migration through K19.2.The expressions of HBx and K19 in HCC are correlated.Positive HBx and K19 suggest that the tumor stage is late and the degree of malignancy is high,which can be used as an indicator for the prognosis of HCC.