| Objective:To value the significance of TGFBR2gene in mediating HepG2cellproliferation by RNA interference technology.Methods:Three kinds of siRNAs targeting TGFBR2gene were designed, synthesized andtransfected into HepG2cells via lipofectamine2000. Real-time PCR was then used todetect the mRNA level of TGFBR2. Among three kinds of siRNAs, only the one withthe most interference efficacy was selected and the correspondent DNA sequence wasinserted into plasmid pEGFP-N3. Then the recombinant plasmid ofsiRNA-pEGFP-N3was transfected into HepG2cell and western blot was used todetect the protein level of TGFBR2. After cultured with5ng/mL or10ng/mL TGF-β1,HepG2cells were detected by MTT assay on their proliferation to select the optimumconcentration of TGF-β1. Then, TGF-β1was used to stimulate HepG2cells with or without siRNA interference and proliferation of HepG2cells was observed.Results:Among these three siRNAs, siRN-1appeared to be the most effective.Compared with blank group, HepG2cells could be inhibited by both5ng/mL and10ng/mL TGF-β1(P<0.05), but there is no significant difference between the lattertwo groups (P>0.05). After stimulated by5ng/mL TGF-β1, proliferation of HepG2cells showed a marked increase in siRNA-1group compared with blank andsiRNA-NC groups (P<0.05). For all that, the proliferation rate was still lower thanthat in normal HepG2cell group without TGF-β1stimulation.Conclusion:By silencing TGFBR2gene, inhibition of TGF-β1signaling pathway to HepG2cells could be decreased, thereby enhancing the cell proliferation. |
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