| Objective:To find tumor related genes by location and function candidate methods and subsequently validate them are usually used strategies now. To filtrate high expression of CDH17 in hepatoma carcinoma cells and investigate the effects of CDH17(liver-intestine cadherin) RNAi in high hepatoma expression carcinoma cell of Biological effect. Based on these results, the present study is to identify the expression of CDH17 gene and explore its function and mechanism of dysregulation, which should lead to discover a new gene related to hepatoma carcinoma. Method: The expression levels of CDH 17 mRNA and protein in hepatoma carcinoma cells(HepG2、MHCC-97H、Huh-7、SMMC-7721) were detected by RT-PCR and Western blotting to filtrate high expression of CDH17 in hepatoma carcinoma cells, with Lipofectamine and plasmid to transfected high expression of CDH17 in hepatoma carcinoma cells. The expression levels of CDH 17 mRNA and protein in cells transfected with CDH17-RNAi were detected by RT-PCR and Western blotting, respectively. The proliferative activity of hepatoma carcinoma cells was detected by CCK-8 method. The migration and invasive capabilities of hepatoma carcinoma cells were detected by scratch-wound migration assay and Transwell invasion assay.Results: The results of RT-PCR and Western blotting revealed that the expression levels of CDH 17 mRNA and protein in MHCC-97 H cells is signifeicantly higher than HepG2、、Huh-7、SMMC-7721 cells.The results of RT-PCR and Western blotting revealed that the expression levels of CDH17 mRNA and protein in MHCC-97 H cells transfected with CDH17-RNAi is low than control group. CDH17 gene was transfected to MHCC-97 H cell line by liposome and G418. In Vitro, MHCC-97 H cells with upregulation of CDH17 growed more slowly than parental cells, and the numbers of colony formation and abilities of migration and invision were significantly decreased in transfected cells.Flow cytometry showed that CDH17 upregulation made more cell apoptosis.but not influenced cell cycle. The proliferation of CDH17-RNAi MHCC-97 H cells was decreased significantly due to CDH17 knock down(P < 0.05). The invasive and migration capabilities of MHCC-97 H cells transfected with CDH17-RNAi were also signifi reduced(P < 0.05). Conclusion: Knock down of CDH 17 may play an important role in the proliferation, invasion and migration of human hepatoma carcinoma cells and it can promote the oncogenesis and progression of human hepatoma carcinoma. |
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