Clinicopathologic Study On MiR-10b, RHOC And Its Related Genes In Hepatocellular Carcionoma And Their Relation With Invasion And Metastasis

Object To evaluate the invasion and metastasis caused by miR-10b,RHOC and related genes in hepatocellular carcionoma(HCC).Materials and methods The clinical and pathological characteristics of 375 cases of HCC were analysised,which were collected from The general hospital of the People's Liberation Army between 2000 and 2005.Of which 185 case's follow up information had been obtained.And 19 fresh HCC specimens and corresponding pericarcinomatous liver tissues were obtained by surgical resection in The general hospital of the People's Liberation Army between 2007 and 2008. Western blotting was used to investigate the expression of RHOC protein.80 cases with follow up information were examined by immunohistochemically staining to investigate the expression of cyclin D1,P21WAF1,p53,ki-67 and GPC3 genes.The relationship between the genes expression and clinicopathologic characteristics of HCC were evaluated,the clinicopathologic factors and the genes were analyzed by Kaplan-Meier and multivariate COX regression analysis. Of the 19 fresh HCC specimens and corresponding pericarcinomatous liver tissues, Real time quantitate PCR was performed to investigate the expression of miR-10b and RHOC mRNA.Results 1.These 375 patients had a median age of 50.57±10.46(range,19-72) with 87.4%HBsAg positive and 4.3%anti-HCV antibody positive;The apparent peak incidence age was 40～60 years old,and the ratio of male to female was 10.7:1;The 3 and 5 year postoperational survival rate were 52%and 38%;The tumour numbers(p=0.000),tumor size(p=0.025),histological pattern (p=0.000),nuclear features(p=0.000),differentiation(p=0.001) and vascular invasion(p=0.000) were significantly correlated with prognosis.The postoperational survival time of thin trabeculae pattern,compact pattern and pseudoglandular pattern were significantly longer than that of thick trabeculae, scirrhous pattern,and solid pattern(p＜0.009);the postoperational survival time of 1 and 2 grade based on nuclear features were significantly longer than that of 3 and 4 grades(p=0.000);The small cell variant,osteoclast-like giant cell variant, and spindle cell variant were mainly composed of thick trabeculae pattern and solid pattern,which were significantly different from that of clear cell variant.(p≤0.006).2.RHOC protein expression was higher in HCC than pericarcinomatous liver tissues in 9/19 cases,and its positive expression was correlated with vien invasion(p=0.020).RHOC expression was positively correlated with cyclin D1 (P=0.023).Increased expression of cyclin D1 was significantly correlated with increasing nuclear grade(p=0.041);High expression of p21^WAF1 and p53 were significantly correlated with poor tumor differentiation(p=0.032,p=0.031) and vascular invasion(p=0.036,p=0.011);High expression of ki67 was significantly correlated with poor tumor differentiation(p=0.004),increasing nuclear grade(p=0.045) and vascular invasion(p=0.001).Expression of GPC3 was positively correlated with multi-tumour numbers(p=0.038) and poor tumor differentiation(p=0.013).3.On COX regression analysis,tumour size(p=0.042),tuomr numbers (p=0.004) and vascular invasion(p=0.000) were independent predictors of overall survival.4.miR-10b and RHOC mRNA expression levels in HCC were higher than that of in pericarcinomatous liver tissues,but lower expression of miR-10b was correlated with vien invasion(p=0.020),contrastly,higher expression of RHOC mRNA was correlated with vien invasion(P=0.041).But the significantly correlation was not found between the expression of miR-10b and RHOC mRNA(p＞0.05).Conclusion 1.As well as tumor size,tumor numbers,tumor differentiation and vascular invasion of HCC,The histological pattern and nuclear features have prognostic significance. 2.miR-10b,RHOC,cyclin D1,ki67 and GPC3 associate with aggressive of HCC;RHOC may also regulate cell cycle;Cyclin D1 and ki67 have prognostic significance;GPC3 can be a tumour marker in HCC;The expression of miR-10b and RHOC mRNA has no significantly correlation.3.On COX regression analysis,tumour size,tuomur numbers and vascular invasion were independent predictors of overall survival.