The Expression And Significance Of RBBP4 In Non-alcoholic Fatty Liver Disease-related Liver Cancer

Objective: With the prevalence of metabolic diseases such as obesity and diabetes,nonalcoholic fatty liver disease(NAFLD)has become one of the major chronic liver diseases in our country,and its related complications have also increased.The discovery of NAFLD-related liver cancer is closely related to obesity and metabolic factors.Using bioinformatics technology to find the differential protein expression in cancer tissues and adjacent tissues in NAFLD-HCC cases,the potential molecular markers retinoblastoma Binding Protein 4(RBBP4)were screened,its expression in NAFLD-HCC cancer tissues and para-cancerous tissues was verified by immunohistochemistry technology,and its possible significance was analyzed.And through animal experiments,a rapid NAFLD-HCC model was established to simulate the human NAFLD-HCC disease process,and to further verify the expression of RBBP4 in cancer tissues and adjacent tissues.Methods: 1.Data collection: Use bioinformatics technology to screen out eligible HCC cases data in The Cancer Genome Atlas(TCGA)database,perform differential gene analysis,and obtain differential genes.And screen out potential molecular markers,analyze the relationship between the differentially expressed target gene and patient staging,age,metastasis,and prognosis.2.Clinical experiment: Collect eligible NAFLD-HCC patients,obtain pathological slices of cancer tissues and adjacent tissues,and observe whether there are difference in the expression of RBBP4 in cancer tissues and adjacent tissues through immunohistochemical methods;Methods to verify the expression of RBBP4 in hepatitis B-related liver cancer,and analyze whether there is any difference with the expression of NAFLD-HCC.3.Animal experiment: Male mice with single gene apolipoprotein E knockout were used as experimental subjects,and they were treated with streptozotocin and high-fat and high-cholesterol diet to obtain a simulation Human NAFLD-HCC rapid model.Obtain liver specimens of NAFLD-HCC mouse model,perform HE staining,and observe liver steatosis,dysplasia and HCC under light microscope.Immunohistochemical method was used to detect the expression of RBBP4 in tumor tissues and surrounding tissues in NAFLD-HCC mouse model.Results: 1.Using bioinformatics technology,374 cases of LIHC were screened out in the TCGA database,and it was found that the expression of RBBP4 and lipid production genes in cancer tissues was stronger than that in normal tissues(P<0.05).At the same time,in 50 cases of LIHC,the expression intensity of RBBP4 in cancer tissues was higher than that in paired adjacent tissues(P<0.05).The expression level of RBBP4 protein was not significantly different from the patient’s age,TNM stage(P>0.05).The overall survival and median survival of liver cancer patients with high RBBP4 expression were shorter than those with low RBBP4 expression(P<0.05).2.The immunohistochemical method found that the expression intensity of RBBP4 in the cancer tissues of 2 NAFLD-HCC patients was higher than that in the adjacent tissues.This phenomenon also existed in 10 cases of hepatitis B-related liver cancer,and it was statistically significant(P<0.05).3.This experiment confirmed that STZ injection combined with high-fat and high-cholesterol diet can induce NAFLD-HCC model.4.In the mouse NAFLD-HCC model,the immunohistochemical method proved that the expression intensity of RBBP4 in cancer tissues was higher than that in adjacent tissues,and the results were statistically significant(P<0.05).Conclusion: 1.The expression intensity of RBBP4 has no significant relationship with LIHC patients’ age,TNM staging.2.LIHC patients with high expression of RBBP4 have a poor prognosis for patients with low expression.3.High-fat and high-cholesterol diet can induce NAFLD-HCC model in mice,proving that dietary factors and obesity are of great significance in NAFLD-HCC.4.In human and animal models,the expression intensity of RBBP4 in cancer tissues is higher than that in adjacent tissues,suggesting that RBBP4 may be a potential tumor-promoting gene for the progression of NAFLD to HCC.