School of Food Science and Engineering, Harbin Institute of Technology, Harbin, People's Republic of China.
Cancer Biother Radiopharm. 2010 Oct;25(5):529-37. doi: 10.1089/cbr.2010.0768.
The β-catenin gene is a critical component of Wnt signaling pathway. Aberrant activation of Wnt/β-catenin signaling and subsequent upregulation of β-catenin is related to enhancing cell proliferation and developing colon polyps and colon cancer. In the present study, the effect of β-catenin knockdown on the growth and survival of the human colon cancer cell line HT-29 was investigated in vitro. The effect of knockdown of β-catenin on cell proliferation was investigated by MTT assay and colony formation. The cell cycle distribution was investigated by flow cytometry. Apoptosis was measured by nuclear staining and flow cytometry. The change of β-catenin and related proteins were determined by western blotting and immunofluorescence. The results showed that small interfering RNA directed against β-catenin markedly inhibited the expression and nuclear translocation of β-catenin and decreased the expression of known target genes such as cyclin D1 and c-myc; HT-29 cell proliferation was inhibited as indicated by growth reduction, cell cycle arrest in G0/G1 phase, and induction of apoptosis; and the inhibition of cell growth may be associated with switching off cyclin D1 and c-myc expression by small interfering RNA targeted against β-catenin in colon cancer HT-29 cells.
β-连环蛋白基因是 Wnt 信号通路的关键组成部分。Wnt/β-连环蛋白信号的异常激活以及随后的β-连环蛋白上调与增强细胞增殖以及发展结肠息肉和结肠癌有关。在本研究中，体外研究了β-连环蛋白敲低对人结肠癌细胞系 HT-29 生长和存活的影响。通过 MTT 分析和集落形成实验研究了β-连环蛋白敲低对细胞增殖的影响。通过流式细胞术研究细胞周期分布。通过核染色和流式细胞术测量细胞凋亡。通过 Western blot 和免疫荧光测定β-连环蛋白和相关蛋白的变化。结果表明，针对β-连环蛋白的小干扰 RNA 显著抑制了β-连环蛋白的表达和核易位，并降低了已知靶基因如细胞周期蛋白 D1 和 c-myc 的表达；HT-29 细胞增殖受到抑制，表现为生长减少、G0/G1 期细胞周期停滞和凋亡诱导；并且抑制细胞生长可能与通过针对β-连环蛋白的小干扰 RNA 关闭结肠癌细胞 HT-29 中的细胞周期蛋白 D1 和 c-myc 表达有关。
