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Study Of The ARHGEF-7 Hepatocellular Carcinoma Of The Nude Mice

Posted on:2017-12-03Degree:MasterType:Thesis
Country:ChinaCandidate:D Q QinFull Text:PDF
GTID:2334330491459208Subject:Surgery
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Objective:To elucidate the role of an oncogene gene ARHGEF-7 in tumorigenesis including proliferation, invasion and migration of HCC. Further more, to explore the function of ARHGEF-7gene mechanism in HCC development, so as to provide new theoretical basis for the HCC diagnosis and treatment.Methods: RNA-interfered lentiviral vector was constructed according to the targe tgene sequence ARHGEF 7.Lentivirus with ARHGEF 7 interference sequence was used to infect human HCCLM3 cells, meanwhile. Nude mice models with HCCLM3 cells transfected by lentiviral vectors were established, Then,take the subcutaneous tumor construction in- situ cultivation of the liver tumor model in nude mice. Through the study of animal model and comprehend the effect of ARHGEF 7 in HCC was investigated.Results:1)The lentiviral vectors,HCCLM3 cell lines were successfully constructed,in which ARHGEF-7genes were down regulated. which was transfected with lentiviral vector, was subcutaneously injected in nude mice. We use the H-E dye detection of ARHGEF7 expression in skin primary malignant tumor to evaluate the efficiency of infection in the body, The results are displayed positive expression in the experimental group and control group in the model.2). Construction of subcutaneous implant tumor models, Application of immunohistochemical detection of ARHGEF7 are negative expression in the experimental group primary malignant tumor and the control group positive expression. Further on the tumor size, growth speed and weight analysis and found subcutaneous tumors in the control group was significantly greater than the experimental group. In the established mice models,lentiviral vector with ARHGEF-7 interference effectively reduced the expression of ARHGEF-7 protein in the screened cells and effectively inhibited the growth of transplantation tumor in nude mice model.3). By building implanted model of the liver tumor in nude mice. Next,We take the lung tissue slices of nude mice to counting the number of pulmonary metastases tumor.The results suggest that inhibition of ARHGEF7 expression in vivo can inhibit lung Metastasis of hepatocellular carcinoma. Further comparison of the experimental group and control group of the nude mouse model of primary tumor size of the liver. we using the immunohistochemical detection of ARHGEF7 protein expression levels of the Metastases in the liver, The results prompted that the control group expression clearly higher than the test group in tumor tissue. This tip that inhibit the expression of ARHGEF-7 can inhibiting growth and metastasis of liver cancer.Conclusion: Reduced ARHGEF-7 expression in HCCLM3 cells, then transplanted into nude mice of the under derma and establishment of model of orthotopic tumor of the liver can cause slow the growth of tumors and inhibition metastasis. Verify the tumorigenicity of ARHGEF-7.
Keywords/Search Tags:hepatocellular carcinoma, ARHGEF-7, proliferation, invasion, metastasis
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