Significance of siRNA-mediated TGFBR2 gene silencing on HepG2 cell proliferation
Li Cheng
Abstract
Li Cheng
Abstract
Objective To value the significance of TGFBR2 gene in mediating HepG2 cell proliferation by RNA interference technology. Methods Three kinds of siRNAs targeting TGFBR2 gene were designed, synthesized and transfected into HepG2 cells via lipofetamine2000. Among three kinds of siRNAs, only the one with the most interference efficacy was selected and the correspondent DNA sequence was inserted into plasmid pEGFP-N3. Then the recombinant plasmid of siRNA-pEGFP-N3 was transfected into HepG2 cell and western blot was used to detect the protein level of TGFBR2. Then, TGF-β1 was used to stimulate HepG2 cells with or without siRNA interference and proliferation of HepG2 cells was observed. Results Among these three siRNAs, siRN-1 appeared to be the most effective. After stimulated by 5ng / mL TGF-β1, proliferation of HepG2 cells showed a marked increase in siRNA-1 group compared with blank and siRNA-NC groups(P 0.05). For all that, the proliferation rate was still lower than that in normal HepG2 cell group without TGF-β1 stimulation. Conclusion By silencing TGFBR2 gene,inhibition of TGF-β1 signaling pathway to HepG2 cells could be decreased, thereby enhancing the cell proliferation.
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Objective To value the significance of TGFBR2 gene in mediating HepG2 cell proliferation by RNA interference technology. Methods Three kinds of siRNAs targeting TGFBR2 gene were designed, synthesized and transfected into HepG2 cells via lipofetamine2000. Among three kinds of siRNAs, only the one with the most interference efficacy was selected and the correspondent DNA sequence was inserted into plasmid pEGFP-N3. Then the recombinant plasmid of siRNA-pEGFP-N3 was transfected into HepG2 cell and western blot was used to detect the protein level of TGFBR2. Then, TGF-β1 was used to stimulate HepG2 cells with or without siRNA interference and proliferation of HepG2 cells was observed. Results Among these three siRNAs, siRN-1 appeared to be the most effective. After stimulated by 5ng / mL TGF-β1, proliferation of HepG2 cells showed a marked increase in siRNA-1 group compared with blank and siRNA-NC groups(P 0.05). For all that, the proliferation rate was still lower than that in normal HepG2 cell group without TGF-β1 stimulation. Conclusion By silencing TGFBR2 gene,inhibition of TGF-β1 signaling pathway to HepG2 cells could be decreased, thereby enhancing the cell proliferation.
Key concepts: Transfection, Small interfering RNA, RNA interference, Gene silencing, Cell growth, Molecular biology, Cell, Gene