Li Xiaohuan,Zhu Rong,Tian Yin,Wang Rongquan.Interference of Asc12 expression in colon cancer epithelial cells leads to their differentiation into phenotype of goblet cells[J].J Amry Med Univ (J Third Mil Med Univ),2012,34(19):1936-1939.
Li Xiaohuan,Zhu Rong,Tian Yin,Wang Rongquan.Interference of Asc12 expression in colon cancer epithelial cells leads to their differentiation into phenotype of goblet cells[J].J Amry Med Univ (J Third Mil Med Univ),2012,34(19):1936-1939.DOI:
Objective To determine the effect of transcription factor Ascl2 on differentiation of colon cancer cells. Methods Colon cancer HT-29 and LS174T cells were transfected with shRNA-Ascl2 interference plasmid and shRNA-control plasmid
respectively. A stably-transfected cell line model was established by screening G418. Effect of shRNA-Ascl2 interference plasmid on expression of goblet cell markers
Muc2 and TFF3 mRNA and protein expression
was detected by RT-PCR and Western blot analysis. Results The successfully constructed shRNA-Ascl2/EPGF plasmid and shRNA-control/EPGF plasmid were confirmed by sequencing. Stably-transfected shRNA-Ascl2/HT-29
shRNA-Ctr/HT-29
shRNA-Ascl2/LS174T and shRNA-Ctr/LS174T cells were obtained by screening G418. RT-PCR and Western blot analysis displayed that the Ascl2 interference plasmid could effectively inhibit the Ascl2 expression in HT-29 and LS174T cells (P0.01) and the Muc2 and TFF3 mRNA expression levels were significantly higher in HT-29 and LS174T cells than in their controls after Ascl2 interference (P0.05). However
no significant difference was observed in the TFF3 mRNA expression level in LS174T cells and controls after Ascl2 interference (P0.05). Western blot analysis showed that the Muc2 protein expression level was significantly higher in shRNA-Ascl2/HT-29 and shRNA-Ascl2/LS174T cells than in their controls after Ascl2 interference (P0.05). Conclusion Interference of Ascl2 expression in colon cancer HT-29 and LS174T cells leads to their differentiation into phenotype of goblet cells.