| Objectives:The essay study IRF7 gene expression in liver cancer, and analyze the association between its expression and clinical data among the development of liver cancer; by decreasing the expression of IRF7, we observed its effect on biological function of liver cancer cell. So we could learn more about relations the expression of IRF7 gene with liver cancer development, and provide experimental and theoretical basis on the mechanism of liver cancer and targeted therapy. The part one: IRF7 expression and its relationship with clinical data in hepatocellular carcinoma Purpose:We analyze differences of IRF7 expression in tumor and corresponding adjacent tissues, and explore relationship between IRF7 expression and liver cancer. Method:1. Application Western Blot and immunohistochemistry detect the expression of IRF7 among hepatocellular carcinoma and the corresponding cancer tissues.2. The combination of clinical data, analyze the relationship between IRF7 gene expression and clinical features such as gender, age, serum AFP, cirrhosis, tumor size, tumor stage, tumor metastasis. Results:1.Western Blot detected the 40 HCC tissues and corresponding adjacent liver tissue, IRF7 protein levels in 29 patients(72.5%) carcinoma tissue are significantly higher than the corresponding adjacent tissues of cancer, and 11 cases(27.5%) in adjacent tissues of cancer are higher than carcinoma tissues.2. Immunohistochemical detection found that among 60 cases of hepatocellular carcinoma IRF7 high expression was 65.0%(39/60 patients), high expression of cancer adjacent tissues was 38.3%(23/60 patients).The high expression rate of IRF7 in liver cancer tissues was significantly higher than in the adjacent tissues, and statistically significant(P<0.05);3. Analysis of the relationship between IRF7 expression and clinical parameters, we found positively correlation between the expression of IRF7 and tumor size, metastasis, and negatively correlation with degree of differentiation. Conclusion:IRF7 gene expression is relation with invasion of liver cancer, and plays an important role in the development of hepatocellular carcinoma.The part two: IRF7 expression impact hepatoma cell biological function Purpose:We further study that gene expression of IRF7 effect on hepatoma cell proliferation, cell cycle, invasion in cell experiment. Method:1. Method of q RT-PCR and Western Blot detect the expression of the IRF7 target gene in normal liver cell line(HL-7702), and different hepatoma cell lines(Hep G2, Huh-7, Hep3 B, SMMC7721, and MHCCLM3).2. RNA interference specific inhibit the expression of IRF7 gene in hepatoma cells, meanwhile we can use CCK8, flow cytometry, Transwell assay to observe the effects on liver cancer cell proliferation, cell cycle, invasion and metastasis after reducing IRF7 gene expression, respectively. Results:1. q RT-PCR and Western Blot results suggest that, IRF7 mRNA and protein expression in hepatoma cell lines are higher than in normal liver cells; and IRF7 expression in a human hepatoma cell line MHCCLM3 are higher than the others.2. RNA interfere IRF7 expression in MHCCLM3 hepatoma cell lines, by q RT-PCR and Western Blot detected the effect on IRF7 mRNA and protein expression levels after RNA interference, and selected the best series of RNA interference.3. We found to reduce the expression of IRF7 genes, liver cancer cell proliferation and invasion period were significantly inhibited by the detection of CCK8 testing, flow cytometry, Transwell assay Conclusion:The expression of IRF7 gene is higher in hepatoma cells. IRF7 expression can promote liver cancer cell proliferation, cell cycle and invasion. Summary:This essay detected IRF7 expression in HCC tissues, and analyzed the clinical datas of patients with liver cancer. So that we found that the expression of IRF7 is closely related to the occurrence and development of HCC. Meanwhile, cell biology function experiments confirmed IRF7 expression can promote liver cancer cell proliferation, cycle, invasion; further revealed IRF7 gene expression played an significant role in the progression of liver cancer. |
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