Mechanisms Of Hepatitis C Virus Infected Cells By Hypervariable Region1

Objectives HCV (hepatitis C virus, HCV) were the major cause that lead to end-stageliver disease such as cirrhosis and hepatocellular carcinoma.Currently,there were nospecific vaccine and therapeutic drugs targeting HCV.And clarifying the mechanism ofHCV endocytosis makes it possible to treat HCV. HVR1located at the N-terminus ofHCV envelope protein E2was the key area that mediates the binding of protein E2andcell receptor SR-BI. Establishing the model, explore the critical molecules that effect thebinding of HVR1protein and the cell,and furthermore explore its impact on the cellsignaling pathway downstream after binding.Methods Purifyly prokaryotic expressed HVR1protein, HVR1protein,Raji cells andL-02cells were co-incubated. Incubated with FITC-labeled HVR1antibody,and thenidentified with FACScan flow cytometer。Silence the SR-BI gene by small interferingRNA method,assess the silence effects by real-time quantitative PCR(Q-PCR) andwestern-blot,and observe the combination efficiency by FACScan flow cytometer.Western-blot analysed the change of ERK, JNK signaling pathway after HVR1proteinbinded to cells,and the change of JNK signaling pathway after HVR1protein binded toL-02cells with the existence of LDL.Results30min after HVR1protein co-incubated with Raji cells and L-02cells underthe protein concentration of200μg/ml,the combination efficiency was52.4%and58.9%respectively. The combination efficiency between HVR1protein and Raji cell was noobvious change under the condition of LDL,but the efficiency between HVR1proteinand L-02cell was rised by85%. The combination efficiency of HVR1protein and Rajicell,and the combination efficiency of HVR1protein and L-02cell were decreased by63%and66%respectively after the silence of SR-BI gene. HVR1protein could activethe ERK, JNK signaling pathway in L-02cell,but no activation in Raji cell. HVR1protein could obviously suppress the JNK signaling pathway under the condition of LDH.Conclusions SR-BI receptor plays an important role in mediating the combinationprocess of HVR1protein, Raji cell and L-02cell. LDL could enhance the combinationefficiency between HVR1protein and L-02cell after the binding, the ERK, JNKsignaling pathway could be actived LDL but JNK signaling pathway would be suppressed with the existence of LDL. Therefore, during the process of HCV infectingcells,firstly HVR1combined with SR-BI receptor,and adsorbed on the cell surface withthe help of LDL,and then actived the ERK signaling pathway and the JNK signalingpathway,finally leaded to unlimited proliferation,and enhanced the cellular resistance aswell.