Study Of Aurora-B Gene Promote Migration And Invasion Of Hepatocellular Carcinoma And Its Mechanism

Hepatocellular carcinoma is the most common malignancy in the world,there are about 50-100 million new cases every year,among the seventh leading cause of cancer death.In China,hepatocellular carcinoma mortality rate is the second cause in cancer mortality rate.operation excision rate is low,poor prognosis has been plagued in clinical,recurrence and metastasis of hepatocellular carcinoma is a major cause of death in patients.Therefore,to investigate the molecular mechanism of HCC metastasis and recurrence of HCC is a hot research recently.Aurora-B has been confirmed to have relationship with the formation of tumor,its overexpression causes centrosome amplification,polyploidy formation and P53 deletion,most scholars think Aurora B is a cancer gene.It has been found that highly expressed in many cancers,including: colon cancer,thyroid cancer,oral cancer,non small cell lung cancer,breast cancer.At present no invasive research aspects in hepatocellular carcinoma.The study is divided into three parts to elaborate the Aurora-B gene promotes study on hepatocellular carcinoma metastasis and its mechanism.Part I : The expression and significance of Aurora-B protein in hepatocellular carcinoma tissuesObjective: Learn to find evidence of Aurora-B in hepatocellular carcinoma tissue,and lay the foundation for the further study of Aurora-B in hepatocellular carcinoma and its molecular mechanism.Methods: Using immunohistochemical methods to detect the Aurora-B expression in 80 cases of human hepatocellular cancer and 30 cases of carcinoma adjacent tissues,analysis of Aurora-B expression in HCC relationship with age,sex,clinical stage and prognosis.Results: the Aurora-positive B staining was localized in the nucleus of hepato-cellular carcinoma cells,in 80 cases of HCC tissue samples,55 cases of Aurora-B were positive,the positive rate was 68.8%;In 30 cases of hepatocellular carcinoma tissues,2 cases of Aurora-B positive,the positive rate was 6.67%(P< 0.001).Aurora-B positive expression rates had significant relationship with stage,tumor size,portal vein tumor thrombus(P < 0.05),Aurora-B positive rate expression had no significant relationship with age,gender,level of AFP(P > 0.05).Aurora-B is highly expressed in HCC,the later clinical stage,tumor size,portal vein tumor thrombus in patients with hepatocellular carcinoma specimens in Aurora-B positive expression rates higher than the early clinical stage,tumor size,no vascular invasion HCC.Conclusions: Over expression of Aurora-B in hepatocellular carcinoma related with its prognosis and recurrence,suggesting that Aurora-B is a potential indicator to the prognosis of hepatocellular carcinoma.Part II: EffectsIof micro RNA interference in Aurora-B gene on invasion and metastasis of hepatocarcinoma cell line HepG2.Objective: To investigate the effects of Micro RNA interference in Aurora-B gene on invasion and metastasis of hepatocarcinoma cell line HepG2.Methods: Using hepatocarcinoma cell line HepG2 as research object,the application of artificial constructs targeting Aurora-B gene Micro RNA recombinant plasmid interfering transfected cells.By wound healing and Transwell invasion assay in vitro,detecting the expression level of Aurora-B inhibition effect on invasion and metastasis of hepatocellular carcinoma cells with high expression of Aurora-B,reverse demonstration Aurora-B inhibition promote to cancer cell invasion and metastasis.Results: The study successfully constructed the effective target expression plasmid to the Aurora-B gene of Micro RNA,AURKB-RNAi(27193-1)transfected cell migrationrate was significantly lower than that of empty vector transfected group(P < 0.05),targeting recombinant plasmid of Micro RNA AURKBRNAi Aurora-B(27193-1)transfected group was significantly lower than the invasion of tumor cells transfected with empty vector group(P<0.05),Conclusion: The study confirmed downregulated the expression level of Aurora-B gene in hepatocellular carcinoma cells can inhibit the migration,invasion of hepatocellular carcinoma cells in vitro.Part III : Micro RNA interference of Aurora-B inhibit PI3K/AKT/NF-κ beta pathway in human hepatoma cell migration and invasionObjective: Micro RNA interference of Aurora-B result in the inhibition of PI3K/AKT/NF-κβ pathway can inhibit cell migration invasion.Methods: In the study,Using hepatocellular carcinoma cell line HepG2 as the research object,the application of gene interference,Western blot and other molecular biological methods,by wound healing and Transwell to study molecular mechanism of Aurora-B inhibition of HER2/PI3K/Akt/NF-κβ signaling pathway on invasion and metastasis of hepatocellular carcinoma.Results: Micro RNA interference of Aurora-B downregulation of AKT/ NFkappa /MMPs signaling pathway related protein expression level,compare to empty vectortransfected group AKT,NF-κ B,MMP2,MMP9 protein decreased expression of in the HepG2 cell was transfected by Aurora-B gene target Micro RNA recombinant plasmid(p<<0.05).Conclusion: Interfering Aurora-B can down regulate the phosphorylation level of AKT,inhibit NF-κ B nuclear transferred,inhibit the expression and downregulation of MMP2,MMP9,inhibit HepG2 hepatocarcinoma cell migration and invasion.