Effect Of Sileacing ARAF On The Malignant Phenotypes Of Gallbladder Cancer

Objective:To access the expression level of ARAF in gallbladder carcinoma,to elucidate its regulation of the malignant phenotype of gallbladder tumor,and provide a new theoretical basis for clinical treatment of this disease.Methods and Materials:Continuous collection of 12 cases of gallbladder cancer tissues and 12 cases of normal gallbladder tissues confirmed by pathological examination from February to September 2018 in Shaoxing People's Hospital.The expression levels of ARAF in the two groups were detected by RT-qPCR and Western blot.Transfected with ARAF siRNA and siRNA control respectively,two groups of cells were used to confirm the regulation of ARAF on cell proliferation and PCNA expression.The effect of ARAF on its tumorigenic ability was proved by colony formation experiment.The wound healing and transwell assays were applied to verify the impact of silencing ARAF on cell migration and invasion.The effect of abnormal expression of ARAF on the growth of gallbladder carcinoma was studied by establishing an animal model of transplanted tumor.Student's t-test was performed on the experimental data,and statistical differences were compared.Results:The experiment results showed that the expression of ARAF mRNA and A-Raf protein in gallbladder carcinoma tissues were significantly higher than those in non-tumor tissues(P<0.05).Compared with the siRNA control group,ARAF siRNA can effectively reduce the proliferation,colony formation and proliferating cell nuclear antigen protein(PCNA)expression of gallbladder cancer cells(<0.05);after transfected with ARAF siRNA,cell tolerance to gemcitabine and cisplatin was negatively affected(P<0.05);in the wound healing and transwell assays,silencing ARAF resulted in impairment of cell migration and invasion(P<0.05).In addition,it was further confirmed by xenografts formation assay that the tumor volume and weight of the ARAF siRNA group were significantly lower than those of the control group,which was consistent with the in vitro results(P<0.05).Conclusions:ARAF is up-regulated in gallbladder carcinoma as a proto-oncogene.Silencing ARAF inhibits the proliferation,migration and invasion of gallbladder cancer cells,enhances its sensitivity to chemotherapeutic drugs,and inhibits the growth of tumor cells in nude mice.Therefore,ARAF should be regarded as oncogene in GBC and a potential target for clinical treatment.