高级搜索

HNF4A充当肿瘤抑制基因抑制结直肠肿瘤进展的实验

Experiment of HNF4A as a Tumor Suppressor Gene Inhibiting Progression of Colorectal Cancer

  • 摘要:
    目的 探究HNF4A在结直肠肿瘤进展中的作用。
    方法 构建HNF4A过表达或敲除的结直肠癌细胞系,通过实时定量PCR和Western blot检测构建的细胞系HNF4A的表达水平;通过软琼脂及平板克隆形成实验检测细胞系的克隆形成能力,CCK-8法检测细胞的增殖能力;Western blot检测干细胞标志物的表达水平;最后通过Transwell迁移与侵袭实验检测细胞的迁移侵袭能力,R2基因分析平台分析结直肠癌中HNF4A与基质金属蛋白酶MMP2及MMP9表达的相关性并通过Western blot验证。
    结果 过表达HNF4A抑制了结直肠肿瘤细胞的增殖与克隆形成能力及迁移侵袭能力,敲低HNF4A促进了结直肠癌细胞的增殖与克隆能力及迁移侵袭能力,Western blot显示HNF4A抑制了结直肠癌干细胞的标志物CD133、CD44及EpCAM的表达和基质金属蛋白酶MMP2及MMP9的表达,在HNF4A敲低的细胞系中得出了同样的结论。
    结论 HNF4A通过抑制结直肠癌干细胞特征及金属蛋白酶的表达来抑制结直肠肿瘤的进展。

     

    Abstract:
    Objective To investigate the role of HNF4A in the progression of colorectal cancer.
    Methods We constructed colorectal cancer cell lines with HNF4A overexpression or knockdown. The expression level of HNF4A was detected by real-time quantitative PCR and Western blot; the colony forming ability of the cell lines were tested by soft agar and plate clone formation experiments, and the cell proliferation ability were checked by CCK-8 method; the expression levels of stem cell markers were detected by Western blot. The cell migration and invasion abilities were detected by Transwell migration and invasion experiments. The correlation between HNF4A and matrix metalloproteinases MMP2, MMP9 expression in colorectal cancer was analyzed by R2 gene platform and further verified by Western blot.
    Results The overexpression of HNF4A inhibited the proliferation, colony formation, migration and invasion of colorectal tumor cells. The knockdown of HNF4A promoted the proliferation, colony formation, migration and invasion of colorectal cancer cells. HNF4A inhibited the expression of colorectal cancer stem cells markers, such as CD133, CD44 and EpCAM, as well as the expression of MMP2 and MMP9. Moreover, the same conclusion was obtained in HNF4A-knockdown cell lines.
    Conclusion HNF4A inhibits the progression of colorectal cancer by inhibiting the stem-like cell characteristic and the expression of metalloproteinases.

     

/

返回文章
返回