Experimental Study On The Expression Of Livin, Survivin In Hepatocelluar Carcinoma And Which Induced By 5-Fu

Objective:The primary liver cancer is the most common malignant tumors in our country. The death rate of the primary liver cancer was the second in our country which counted by the Health Department. it has the increased uptrend in its incidence , recurring rate and death rate recently. It is the important reason that the invasion and metastasis of HCC are often the failure of treatment and effect the patients life. The invasion and metastasis of HCC is controlled by many special genes, and is a complex process of polygenes and multistages, and comes down to many tumor genes and suppressors, and these mechanisms are still elusive. Inhibitors of apoptosis proteins(IAPs) was a group of apoptosis regulator, which could ihibit cell apoptosis. Livin is the lastly founded Inhibitors of apoptosis protein.The livin gene was identified by IAP homologics sequence by Lin at 2000.It was founded at human fetal kidney cDNA data bank by EST clon which include BIR sequence and named kidney inhibitor of apoptosis protein(KIAP). Vucic founded that livin gene was high expression at human melanoma and named it melanoma inhibitor of apoptosis protein(ML-lAP). Kasof founded this gene by homologics and named it livin. Survivin is the most small IAP.Both livin and survivin have a N-terminal baculovirus lAP repeat(BIR)domain. Livin has a COOH—terminal RING finger domain but survivin has not.The study suggests that livin and survivin do not or only lower expression in normal differated tissure,but it high express in tumor tissure.It suggest that livin and survivin are correlated with tumor growth, invasion, metastasis.The main mechanism which it inhibit apoptosis is interaction with caspase and ihibit caspase's activation. Caspase activated is the key step of apoptosis.This study will investigate the expression of livin,survivin in HepG2 and investigate the change of livin,survivin after tretment with 5-Fu. This study not only furthered to detect the mechanisms of molecule biology of livin,survivin in HCC, but also provided potential candidates for gene therapy of hepatocellular carcinoma.Methods:1.To culture HepG2 cells.2.Low concentration of 5-Fu was added in cultured HepG2 cells.The expression of livin mRNA and survivin mRNA was detected by RT-PCR on 24h and 48h after 5-Fu tretment.3.Low concentration of 5-Fu was added in cultured HepG2 cells.The expression of livin protein and survivin protein was detected by western-blot on 24h and 48h after 5-Fu tretment.Results: 1. The culture of HepG2 was accomplished.2.It was identified that HepG2 express Livin mRNA,Survivin mRNA. Livin mRNA and survivin mRNA 1eve1 was increased after 5-Fu tretment. The comparation of density in the three group, P<0.05 ,the prominent diversities were affirmed among them.3. It was identified that HepG2 express Livin protein , Survivin protein.Livin protein and survivin protein 1eve1 was increased after 5-Fu tretment。The comparation of density in the three group, P<0.05 ,the prominent diversities were affirmed among them.Conclusions:1. It was identified that HepG2 express both livin and survivin.2. Livin and survivin mRNA 1eve1 was increased after 5-Fu tretment.the diversities were prominent. A high expression level of livin and survivin was induced in HepG2 by 5-Fu,which reduced to apoptosis by 5-Fu.