[HBx silencing by shRNA down-regulates MMP-2 expression in human hepatocellular carcinoma cell HepG2.2.15].
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).


CONCLUSIONS
HBx gene RNA interference can inhibit the cell proliferation and down-regulate the expression of MMP-2 in HepG2.2.15 cells.
Department of Biochemistry, Zunyi Medical College, Zunyi 563099, China.
Changfu
C
Li
Department of Bio-Pharmaceutics, Tianjin Vocational College of Bioengineering, Tianjin 300462, China.
Yuepeng
Y
Sun
Tianjin Pharmaceutical Research Institute Company Limited, Tianjin 300000, China.
Lei
L
Geng
Department of Biochemistry, Zunyi Medical College, Zunyi 563099, China.
Fang
F
Fan
D006528:Carcinoma, Hepatocellular / Q000473:pathology*; D049109:Cell Proliferation; D015536:Down-Regulation / Q000235:genetics*; D056945:Hep G2 Cells; D006801:Humans; D008113:Liver Neoplasms / Q000473:pathology*; D020778:Matrix Metalloproteinase 2 / Q000172:deficiency* / Q000235:genetics* / Q000378:metabolism; D009362:Neoplasm Metastasis; D010957:Plasmids / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics*; D015534:Trans-Activators / Q000235:genetics*; D054334:Viral Regulatory and Accessory Proteins
D034741:RNA, Small Interfering; D015534:Trans-Activators; D054334:Viral Regulatory and Accessory Proteins; C057628:hepatitis B virus X protein; D020778:Matrix Metalloproteinase 2