The Expression And Functional Mechanisms Of UMtCK In Hepatocellular Carcinoma

Object: To investigate the diagnosis and prognosis feasibility of ubiquitous mitochondrial creatine kinease(uMtCK)for hepatocellular carcinoma(HCC)and to investigate the molecular mechanisms and genetic role of uMtCK in HCC cell lines.Method: To investigate and analyze the serological expression of uMtCK in patients with HCC,liver cirrhosis and chronic hepatitis B and different stages of HCC patients using Elisa detection.Receiver Operator Characteristic Curve(ROC)and Area Under the Curve(AUC)were used to detect and compare the diagnosis feasability of uMtCK and AFP expression in the serum of HCC patients.The immunohistochemistry method was used to detect and compare the uMtCK expression in the pathological tissues of HCC patients,liver cirrhosis patients,chronic hepatitis B patients to analyze the correlation between uMtCK and the malignant biological characteristics of HCC.The MTT method was used to investigate the proliferation role of HCC cell.The flow cytometry method was used to investigate the apoptosis role of HCC cell.The si-RNA was used to knock down the expression of uMtCK,HCC cell proliferation,migration and invasion were detected through Brd U incorporation assay and transwell method.Results:(1)The serum expression of uMtCK in HCC patients is significantly higher than the serum expression of liver cirrhosis patients and hepatitis B patients(P <0.05);The serum expression of uMtCK in late stage of HCC patients is significantly higher than the serum expression of early stage of HCC patients(P <0.05);The AUC of uMtCK and AFP were similar when comparing the feasability of diagnosis for HCC,(P >0.05).The specificity of AFP is significantly higher than uMtCK(P <0.05).The sensitivity of AFP and uMtCK is similar with each other(P > 0.05);The sensitivity of cooperation diagnosis of AFP and uMtCK is significantly higher than the sensitivity of AFP(78% vs 54%,P <0.05);The expression of uMtCK is significantly decreased after the curative therapy of RFA for HCC patients(P <0.05).(2)The expression of uMtCK is significantly higher in the pathologic tissues of HCC patients than the expression of uMtCK in the pathologic tissues of hepatitis B patients(P <0.05);The expression of uMtCK exists the positive relation with HCC stages and the invasion of portal thrombosis.(3)Western Blot showed the high expression of uMtCK in the cells of Hep G-2 and Huh-7 cell lines(P <0.05);MTT method showed that high expression of uMtCK can proliferate HCC cells;Flow cytometry showed uMtCK-pc DNA3.1(-)transfected cell lines significantly inhibited Hep G-2 and Huh-7 cells apoptosis(P <0.05);The ability of proliferation,migration and invasion were significantly inhibited when the expression of uMtCK is knocked down through si-RNA(P <0.05).Conclusions:uMtCK is over expressed in the serum and the pathological tissue of HCC patients.Over expressed uMtCK may plays the role of proliferating,migarating and inviding of HCC cells,and the role of inhibiting apoptosis of HCC cells.Therefore,uMtCK maybe become an assisting serological bio-marker for the diagnosis and prognosis feasibility of HCC.