Hppcn In Liver Cancer, The Function And Mechanism In The Development Of Study

Hepatocellular carcinoma (HCC) is the most common primary liver cancer and fifth most common cancer in the world.HCC is critical disease to harm health of people. Recent report suggest an increase in the incidence of HCC in the world.In our country, there are about 11 ten thousands people were died because of HCC.HCC not sensitive to systemic chemotherapy ,and have low efficiency to partical liver resection.Now systemic therapy strategy is deficiency to HCC,and in the process of management patient easy to recur and drug fast,At the end ,this result induce to defeated in therapic process.For this reason ,it's the important scienctic question about to people weathy ,that study pathpgenesy of HCC,and develop new therapy stratagy.HPPCn is isolated a pure protein with hepatic stimulatory activity from the weanling calf liver. Bioactivity study to HPPCn suggested that rhHPPCn can specific stimulate the proliferation of primary cultured rat liver cells,human liver tumor cell SMMC7721/HepG2 and L02 cell lines,also can enhance the regeneration of hepatectomized liver.Animal experiment study indicated that rhHPPCn can protect acute liver injury and hepatic fabrosis induced by CCl4.In order to study HPPCn and signalway function in the process of HCC hepatocarcinogenesis and development ,we are carry out some work to this study.Firstly ,we are constructed and preparated that HPPCn liver specific and eukaryotic expression vector ,which have ALB promoter and enhencer suquence.After that ,we are verificated the vector liver specific expression ability in vitro.Result indicted that HPPCn liver specific expression vector can make HPPCn expression in hepatic cell line. HPPCn expression in HEK293,L02,SMMC7721,HepG2 cell is 0.94%,35.48%,58.94%,62.51%.This work is establish fundation of HPPCn liver expression transgene mice to study HPPCn function in hepatocarcinogenesis.Meanwhile ,we are contructed HPPCn interfereced vector pGPU6/GFP/Neo-shRNA,which made HPPCn expression downregulated in liver tumor cell,to investigated influence to HCC development.We are utilizted this vector made HPPCn expression downreguated 76.4%.Because the pGPU6/GFP/Neo-shRNA contained an SV40-driven neomycin selectable marker,which confers resistance to G418 antibiotic,we can selected HPPCn stabilizated interferenced cell clone.Secondly,we are carry out study,which surrounding HPPCn bioactivity to tumor cell and liver stem cell .Our study show that ,rhHPPCn can protect hepatoma carcinoma cell to apoptosis induced by TSA,and can promote hepatoma carcinoma cell proliferation,and up-regulate expression of MCl-1 .We are confirm that rhHPPCn can activate PI3K/AKT,JAK-STAT3 signalways to up-regulation expression of MCl-1.Next,we are interferece the expression of HPPCn by RNAi ,so that make expression of HPPCn down-regulated in hepatoma carcinoma cell.We are confirm that HPPCn interfereing can inhibite hepatoma carcinoma cell proliferation and migration,enhance cell apoptosis rate,degrate hepatoma carcinoma cell cloning-efficiency.Then, we are utilize WB-F344 cell as model to study HPPCn bioactivity to liver stem cell,result show that HPPCn can promote WB-F344 cell proliferation and migration,also can increase cell cloning-efficiency.Finally,we investigated the influecne to tumor development by HPPCn in vivo.We are to use BAL/C nude mice as model,constructed normal HepG2 cell and HepG2/si-HPPCn cell transplantation tumor model.The result suggestion that the tumor of transplanteing HepG2/si-HPPCn cell is smaller than transplanting normal HepG2 cell, growth velocity of HepG2/si-HPPCn cell is step down.Meanwhile ,we are to supply Ad-HPPCn to nude mice of transplanted normal HepG2 cell,the result indicated that Ad-HPPCn can stimulating tumor growth in vivo.In conclusion,our study indicate that HPPCn and its signalway have important function in the process of tumorigenesis and development. Our study result can provide new therapy target.