Inhibition of CD133 by RNA Interference in human colon carcinoma cell line HT-29

OBJECTIVE: To construct an expression plasmid of siRNA against CD133 gene and to investigate its biological behavioral effect on human colon carcinoma cell line HT-29.METHODS: we inhibited the CD133 expression using the siRNA method.Expression levels of CD133 mRNA and protein from HT-29 after transfection were examined by RT-PCR and Western blot respectively.Changes of the cell growth activity in response to transfected plasmid were evaluated by MTT assay.The migration ability of transfected HT-29 cells were measured quantitatively by Transwell migration assays.RESULTS: After CD133 siRNA transfeetion into HT-29 cells,compared with that in control groups,the level mRNA of CD133 in cancer cells transfected with CD133 siRNA was inhibited in a concentration and time dependent manner and the CD133 protein concentration decreased significandy.The growth of HT-29 cells was significantly inhibited than those in control groups(P0.05).CD133 siRNA significandy inhibited invasion of HT-29 cells to matrigel(P0.05).CONCLUSION: Silencing CD133 gene by the siRNA technology can actively suppress the expression of CD133 gene,and then inhibit the growth and invasion of HT-29 cell.