Preliminary Study Of RBM8A Regulating The Resistance Of Hepatocellular Carcinoma To Oxaliplatin

Objective:Previous studies have found that RBM8 A plays an important role in the occurrence and development of malignant tumors and is up-regulated in many malignant tumors.The aim of this study was to compare the expression of RBM8 A in the serum of patients with primary liver cancer,cirrhosis,other malignant tumors and healthy subjects,and the changes of serum RBM8 A expression in patients with primary liver cancer before and after surgery.The potential of RBM8 A as an indicator for the diagnosis of hepatocellular carcinoma;the effect of RBM8 A expression on the apoptosis,proliferation and drug resistance of parental strains and oxaliplatin-resistant hepatocarcinoma cells was investigated by cell assay.Mechanism of action on oxaliplatin resistance.Methods:1.A total of 706 peripheral blood samples were collected,including137 patients with primary liver cancer,33 patients with liver cirrhosis,172 patients with other malignant tumors,and 56 healthy subjects.Further,serumsamples from patients with primary liver cancer after surgery were collected.The levels of RBM8 A in the serum of each group were also measured by double antibody clamp and enzyme-linked immunoassay.2.The OXA-resistant in vitro hepatoma cell model Bel 7404/OXA and MHCC-97H/OXA were established by increasing the concentration of drug in vitro.The morphology of the cells was observed under an inverted phase contrast microscope.3.Hepatocellular carcinoma cell lines were infected with RBM8 A overexpressing lentivirus and RBM8 A to interfere with lentivirus,and a stable transfected cell line model was selected-MHCC-97H-RBM8A-OE,MHCC-97H/OXA-RBM8A-OE,Bel 7404-RBM8A-KD,Bel7404/OXA-RBM8A-KD was verified by Western blot.4.The cell proliferation ability and IC50 of each strain were detected by CCK-8 method;the degree of apoptosis of cells was detected by flow cytometry.5.The expression levels of drug resistance proteins ABCG2,P-GP and MRP1 were detected by Western blot.Results:1.The expression of serum RBM8 A in patients with primary liver cancer was significantly higher than that in cirrhosis group,other malignant tumor group and healthy control group(204.29±224.42 VS.10.55±7.53,3.33±1.71,2.13±2.04,P<0.001);primary After surgical treatment of liver cancer patients,the expression of serum RBM8 A decreased compared with that before treatment(167.93±205.9 VS.204.29±224.42,P<0.001).2.The OXA resistance model Bel7404/OXA,MHCC-97H/OXA was successfully constructed.Under the inverted phase contrast microscope,themorphology of drug-resistant cells was changed from uniform circular or round-like epithelial-like cells to interstitial-like phenotypes of various irregular cell shapes such as long strips,fusiforms,and polygons.Western blot The results indicated that the expression of RBM8 A in drug-resistant cell lines was higher than that of the parental cell line.3.CCK8 results showed that RBM8 A was overexpressed in MHCC-97 H cells in parental or drug-resistant hepatoma cell lines,which promoted cell proliferation,increased IC50 value,and increased tolerance to oxaliplatin;Silencing RBM8 A in BEL 7404 cells,Inhibition of cell proliferation,decreased IC50 value,increased sensitivity to oxaliplatin.4.The results of flow cytometry showed that RBM8 A was overexpressed in MHCC-97 H cells and inhibited apoptosis in parental or drug-resistant hepatoma cell lines.Silencing of RBM8 A in BEL 7404 cells promoted apoptosis.5.Western Blot results showed that in the parental or drug-resistant hepatoma cell line,RBCC8 A cells overexpressed RBM8 A,the expression level of drug resistance-related protein was increased,and RBM8 A was silenced in BEL 7404 cells,and the expression level of drug resistance-related protein was decreased.Conclusions:1.RBM8 A has potential value in the diagnosis and recurrence of primary liver cancer.RBM8A is involved in apoptosis,proliferation and drug resistance of hepatocellular carcinoma cells.Silencing RBM8 A can promote the apoptosis of hepatoma cells,inhibit the proliferation of hepatoma cells,and down-regulate the expression of drug resistance-related proteins in hepatocarcinoma cells.Overexpression of RBM8 A can inhibit the apoptosis of hepatoma cells.Death,promote the proliferation of liver cancer cells,up-regulate the expression of drug resistance-related proteins in liver cancer cells.3,RBM8 A may be involved in the regulation of oxaliplatin resistance in liver cancer.