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  • Title: [HBx silencing by shRNA down-regulates MMP-2 expression in human hepatocellular carcinoma cell HepG2.2.15].
    Author: Li C, Sun Y, Geng L, Fan F.
    Journal: Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi; 2014 Apr; 30(4):371-4. PubMed ID: 24721404.
    Abstract:
    OBJECTIVE: To observe the effects of HBx shRNA on MMP-2 expression in human hepatocellular carcinoma cell HepG2.2.15. METHODS: HepG2.2.15 cells were transfected with psiHBV/X plasmid using Lipofectamine(TM);2000. The silencing efficacy was evaluated by reverse transcription PCR (RT-PCR) detecting the expression of HBx gene. The proliferation of HepG2.2.15 cells was examined by MTT assay. The expression levels of MMP-2 mRNA and protein were assayed by real-time quantitative PCR (qRT-PCR) and Western blotting, respectively. RESULTS: RT-PCR showed that the efficiency of RNA interference of HBx gene was 53.6%. MTT detection revealed that the cell proliferation was inhibited at 24, 48, and 72 hours after the transfection (0.388±0.087, 0.623±0.016, 0.997±0.036) and the differences had statistically significance as compared with the control group (0.436±0.027, 0.731±0.017, 1.105±0.051) (P<0.05). The qRT-PCR and Western blotting showed that the expression of MMP-2 decreased at both mRNA and protein levels, and the differences had statistically significance as compared with the control group (P<0.05). CONCLUSION: HBx gene RNA interference can inhibit the cell proliferation and down-regulate the expression of MMP-2 in HepG2.2.15 cells.
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