| Primary carcinoma of liver is one of the most common malignant tumors,and its morbidity and mortality stand at the forefront of the world.Hepatocirrhosis is considered as one of uppermost causes of death at present,and its morbidity is likely to go up in the future.In recent years,with the continuous improvement of medical technology,we have developed sophisticated diagnostic methods,mature surgery,advanced medical equipment and auxiliary chemotherapy drugs,and even liver transplantation.However,patients still have recurrence and metastasis problems.Failure to make breakthrough progress,the prognosis of patients is not optimistic.Further research on the regulation mechanism of malignant biological behavior of liver cancer will provide a basis for screening prognostic markers and targeted therapeutic targets for liver cancer.The ubiquitination and deubiquitination of proteins are crucial determining factors of all kinds of biological processes,including regulation of cell cycle,DNA repair,apoptosis,antioxidant properties and hypoxia response,all of which directly or indirectly affect the radiosensitivity of tumors and the radiation resistance of cells.Meanwhile,the chaos of the ubiquitination/deubiquitination system has already been reported to be relevant to the anti-radiation of tumors.Therefore,elucidating the ubiquitin-mediated molecular mechanisms will open up a method for tumor treatment strategies and improve the sensitivity of tumor cells to radiotherapy.PRAJA1 is an E2-dependent E3 ubiquitin ligase,which has the ligase activity of ubiquitin-protein and belongs to a class of ubiquitin ligases containing a ring finger structure.Recent studies have shown that PRAJA is highly expressed in brain tissue,including cerebral cortex,cerebellum,frontal lobe,occipital bone,etc.The unusual expression of PRAJA1 is linked with the development of glioma.Besides,it plays a very significant role in the development of bones.Nevertheless,its mechanism in tumors is less studied,and its role in hepatocellular carcinoma remains unclear.Therefore,this study mainly introduces the role of PRAJA1 in liver cancer.The main results are as follows:1 The expression level of PRAJA1 and survival analysis in liver cancer:(1)Immunohistochemical method for liver cancer tissue staining,the results showed that PRAJA1 is highly expressed in liver cancer tissues compared with normal liver tissue;(2)GEPIA database was used to analyze the relationship between the expression of PRAJA1 and the survival of patients with hepatocellular carcinoma.Compared with patients with high expression of PRAJA1,patients with low expression of PRAJA1 had longer survival time.2 The effect of PRAJA1 on proliferation of hepatocellular carcinoma cells: A stable overexpression and knockout of PRAJA1 in hepatocellular carcinoma cell lines were established,and the effect of overexpression and knockout of PRAJA1 on cell proliferation ability was detected by MTS assay and wound-healing assay assay.The results showed that compared with the control group,the proliferation ability of liver cancer cells increased after overexpression of PRAJA1,and the proliferation ability of liver cancer cells decreased after PRAJA1 knocked out.3 The effect of PRAJA1 on the migration of hepatocellular carcinoma cells: The scratch test method was used to detect the effect of over-expression and knockout of PRAJA1 on the migration ability of Hepatocellular carcinoma cells.The results displayed that compared with the control group,the migration ability of hepatocellular carcinoma cells enhanced after overexpression of PRAJA1 and weakened after knockout of PRAJA1.4 The effect of drug treatment on the expression of PRAJA1 in hepatoma cells:(1)Hepatocellular carcinoma cells were treated with sorafenib,melatonin and five traditional Chinese medicines FM-8,FM-9,FM-14,FM-15 and FM-16,and the results showed that the FM-15 with the lowest expression of PRAJA1 after drug treatment was screened out;(2)The expression of PRAJA1 in hepatocellular carcinoma cells was analyzed by time course and dose-dependent analysis.The results showed that the expression of PRAJA1 decreased gradually with the prolongation of FM-15 treatment time,and decreased gradually with the increase of the concentration of FM-15 treatment;(3)Stable overexpression and knockout of PRAJA1 cell lines treated with FM-15 drug were tested for their proliferative capacity by MTS method.The results showed that knockout of PRAJA1 combined treatment with drugs significantly reduced the proliferation of hepatoma cells.5 Effect of PRAJA1 knockout HCC cells on tumorigenic ability of nude mice: the tumorigenic experiment was conducted on nude mice by subcutaneous injection with PRAJA1 knockout cell line,and the results showed that after PRAJA1 knockout,the growth rate of transplanted tumor was slower,and the final volume of transplanted tumor was smaller and the number was fewer.6 Label-free quantitative proteomics analysis: With high throughput data,the differentially expressed proteins in PRAJA1 knockout hepatocellular carcinoma cells were identified by non-labeled quantitative analysis.Pathway of differentially expressed proteins was identified by Pathway significant enrichment analysis to determine the main biochemical metabolic pathways and signal transduction pathways in which differentially expressed proteins participated.In summary,PRAJA1 is highly expressed in hepatocarcinoma tissues,which can promote the proliferation and migration of hepatocellular carcinoma cells.The Chinese medicine FM-15 can inhibit the expression of PRAJA1,and then inhibit the proliferation of hepatocellular carcinoma cells.The knockout of PRAJA1 can inhibit the tumorigenic ability of hepatocellular carcinoma cells in vivo.That is,PRAJA1 acts as an oncogenic factor in liver cancer. |
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