Of ShRNA-AFP Mitomycin Nude Mice Implanted Liver Cancer

[Objective]Research the effect of RNA interference targeting AFP combined MCC on the therapeutic efficacy on the tumor in nude mice and inquire into the mechanism of apoptosis and resistance in hepatocellular carcinoma,attempting to provide a new target and a new idea for the gene therapy to HCC.[Methods]1. Human hepatocellular carcinoma Huh7cells were cultured in vitro by cell strain subculturing technique.2. Establishment of stable subcutaneous hepatocarcinoma model:non transfected Huh7cells are inoculated into the nude mouse.3.Pick out30excellent nude mouse models and randomly divided them into5groups. A,B, C,D,E,4. Injection of gene therapy or drug everyday, tumor diameter and short diameter were measured at3,6,9,15days, calculate the tumor volume, draw the tumor volume-time growth curve.5. Expression of AFPmRNA and Bcl2mRNA Were detected by reverse transcription PCR(RT. PCR).6. Detect of AFP and Bcl2protein expression by immunohistochemistry.[Results]1. The differences of the gross tumor volume between five groups were significant, group D is smaller than other four groups(P<O.05).2. Results of RT. PCR:the level of Bcl2mRNA in group D decreased significantly compare with other groups (P<O.05).3.Results of SP: the level of AFP and Bcl2protein in group D decreased significantly compare with other groups(P<O.05).[Conclusions]1. The treatment of AFP-shRNA combined MCC can significantly inhibit tumor growth, and also inhibit expression of AFPmRNA and Bcl2mRNA and expression of AFP protein, Bcl2 protein.2. The results of the experiment offers a new theory for gene therapy to HCC, and provide a new target and pathway for gene therapy of tumor.