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.
ABSTRACT
Objective: To screen high-affinity short peptides of the transforming growth factor beta typeⅡ(TGF-? Ⅱ) receptor from the phage display 12-mer peptide library.Methods: The recombinant solube human TGF-? Ⅱ receptor was used as the target to screen its affinity short peptides from the phage display 12-mer peptide library.The positive phage clones were obtained by 3 rounds of biopanning,followed by single stranded DNA extraction and sequence analysis.Results: Four nucleotide sequences were obtained from 10 positive phage clones with high-affinity TGF-? Ⅱ receptor.Neither conservative nor homologous sequence was found by sequence analysis compared with the transforming growth factor beta 1.Conclusion: Phage short peptides have been screened from the phage display library,which helps pave the ground for further researches on affinity short peptides and their role in inhibiting hepatic fibrosis.