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[HBx silencing by shRNA down-regulates MMP-2 expression in human hepatocellular carcinoma cell HepG2.2.15]
Changfu Li1, Yuepeng Sun2, Lei Geng3
1Department of Biochemistry, Zunyi Medical College, Zunyi 563099, China.
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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