CTNNB1基因沉默抑制结肠癌SW480细胞生长的研究

目的探讨人结肠癌SW480细胞的CTNNB1基因沉默效应和对细胞生长的抑制作用。方法构建靶向CTNNB1的shRNA载体质粒,然后转染入结肠癌SW480细胞。用RT-PCR和Western Blot方法检测CTNNB1的mRNA和蛋白的表达情况。MTT实验评价转染后各组细胞的增殖情况。结果靶向CTNNB1的shRNA能够明显下调CTNNB1mRNA和蛋白质表达(P0.05),其抑制率分别是38.29%和42.86%。MTT实验提示转染了特异性CTNNB1shRNA的CTN实验组细胞在转染后呈现随着时间延长而进行性生长抑制的现象。在转染后72h,CTN组的细胞存活率为43.7%,与空白对照组相比有显著性降低。结论靶向CTNNB1的特异性shRNA对结肠癌SW480细胞具有下调CTNNB1基因表达,并显著抑制细胞增殖的效应。

CTNNB1 gene silence effect inhibit the growth of human colon cancer SW480 cells

Objective To observe the inhibitory effect of CTNNB1 gene expression and cell proliferation of the human colon cancer cell line SW480.Methods The shRNA plasmid vector against CTNNB1 was constructed and transfected into SW480 cells with LipofectamineTM2000.The down-regulations of CTNNB1 expressions were detected by RT-PCR and western blot analysis.The cell proliferation inhibitions were determined by MTT assay.Results The shRNA vectors targeted against CTNNB1 were successfully constructed and efficiently suppressed the expression of CTNNB1 mRNA and protein(P<0.05).The expression inhibition rates were 38.29% and 42.86% at the mRNA and protein level respectively.The MTT assay indicated that the specific shRNA resulted in significant inhibition of cell growth on the culture plates on a time-dependent manner.At 72 h post-transfection,the cell viability of CTN group was 43.7%,which was of statistical significance when compared with that of blank control group(P<0.05).Conclusion The CTNNB1 gene silence effect could down-regulate the expression of CTNNB1 gene and inhibit the growth of SW480 cells.