The Expression Of ARID1A Related With Poor Prognosis In Hepatocellular Carcinoma And Its Mechanism

Hepatocellular carcinoma（HCC）is one of the highest incidence of malignant tumorsin China, Although much progress has been made in HCC research, but the invasion andmetastasis are the leading cause for the HCC death. Untill now, tumor resection and livertransplantation are the most effective and standard therapies for HCC, Therefore，it isessential to explore the underlying mechanisms of recurrence and metastasis of HCC tofind effective ways to improve the survival rate of HCC.Tumor development, invasion and metastasis is a complex and dynamic processes.From the perspective of molecular biology, the HCC is a genetic disease. Because ofcertain chromosomes, DNA damage causes the mutation results in regulating genes,including oncogenes, tumor suppressor genes, apoptosis genes, cell cycle, and maintainthe stability of the cell genome gene is a multi-stage process of gradual evolution.Chromatin remodeling complexes SWI/SNF AT-rich interactive domain1A (ARID1A) isan important gene in cancer research in the past two years. It is an important tumorsuppressor gene and closely associated with cell proliferation.ARID1A play an importantrole in tumor development, metastasis and recurrence of the process.There is no people had to reports of the impact in HCC. In the present study, wediscussed the impact of ARID1A on biological behaviors of HCC. We investigated theexpression of ARID1A in HCC Cell lines with different metastatic potential. Tissuemicroarray for240HCC patients showed decreased expression of ARID1A in tumor cellsand study with the development and prognosis in HCC. By in vivo and vitro experiment，we observed the impact of ARID1A expression on proliferation, invasion, tumor growthand metastasis. We hope to provide a new therapeutic target for the prevention andtreatment of HCC. Part I The expression of CD88in hepatocellular carcinomaand HCC cell linesObjective: We focused on the expression of CCRL1in cancer and peri-cancer tissuesand the relations between CCRL1expression and tumorigenesis.Methods: We investigated the expression of ARID1A in HCC Cell lines withdifferent metastatic potential. Performed in cancer and peri-cancer tissue of40patientswho underwent HCC resection.Result: We found that: ARID1A protein level detected by western blot wassignificantly lower in HCC than those in their adjacent nontumorous tissues（P<0.05）. theARID1A mRNA level Was significantly lower in HCC than those in their adjacentnontumorous tissues. The experiment in HCC cell lines, We found that: The level ofARID1A protein and mRNA in HCCLM3and MHCC-97H with high metastasis potentialwas lower than in Hep-G2and Hep-3B with low metastasis potential.Conclusion: ARID1A play important role with the development in HCCPart II Correlation and clinical significance of expressionof ARID1A in hepatocellular carcinomaObjective: To investigate the correlation and clinical significance of ARID1A withHCC.Methods: immunohistochemical staining was performed in tumor and peri-tumortissue of240patients who underwent HCC resection, then the clinical significance ofexpression of ARID1A was analyzed by SPSS16.0.Result: Overexpression ARID1A was found to correlate significantly with high serumAFP level（P=0.031）, sex（P=0.036）, vascular invasion（P<0.0001）, Child-Pugh score（P=0.607). and BCLC stage（P=0.043）.5-year overall survival and disease-free survivalrate in the group of ARID1A-positive was significantly lowerthan that in the group ofARID1A-negative group(p<0.03). Multivariate analysis showed that serum AFP>20ng/ml, liver cirrhosis, Tumor size>5cm and ARID1A positive expression were independentprognostic factors for OS; serum AFP<20ng/ml, liver cirrhosis, Tumor size<5cm andpositive expression of ARID1A were independent prognostic factors for DFS.Conclusion: The low expression of ARID1A can be a new marker in predicting theprognosis of HCCPart III The impact on HCC biological characteristicsby regulation of ARID1A expressionObjective: Explore the impact of ARID1A on tumor growth, invasion, migration,apoptosisand metastasis in vivo and in vitro experiments.Methods: Using lentivirus vector and TALE-VP64, we transfected shARID1Aknock-down and overexpression plasmids in Hep-3B and HCCLM3cell lines, toconstructed stable cell lines of Hep-3B-shARID1A and HCCLM3-ARID1A. theexpression of ARID1A was determined by qRT-PCR, western blot.The ability ofproliferation and invasion and migration was tested by MTT and transwell in the stabletransfection HCC cell and parental cell lines,The ability of apoptosis was tested by Flowcytometry. The human liver cancer mode was established with Hep-3B and HCCLM3cellsin athymic nude mouse. the gross tumor volumes were measured dynamically.Immunohistochemistry for the detection of tumer growth-related factor expression: PTEN，PIK3CA,p53，MMP9，VEGF.Result: In vitro experiments, our results showed that: overexpression of ARID1Ainhibited the proliferation, migration and invasion of HCC cells. On the contrary,down-regulation of ARID1A caused the intensive proliferation, invasion, and migration ofHCC cells.HCC cell lines with high ARID1A expression increase the mRNA expression ofPTEN,p53,PIK3CA and decrease the mRNA expression of MMP9,VEGF.In vivo, thetumor mass in the low expression group higher than the high expression group.Conclusion: ARID1A can inhibit the proliferation, migration and invasion of theHCC cell lins,overexpression ARID1A can increase the expression of PTEN,p53,PIK3CA and decrease the expression of MMP9,VEGF.