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Interference of Ascl2 expression in colon cancer epithelial cells leads to their differentiation into phenotype of goblet cells
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- Author:
- No author available
- Journal Title:
- Acta Academiae Medicinae Militaris Tertiae
- Issue:
- 19
- DOI:
- No doi available
- Key Word:
- Ascl2;结肠肿瘤;肿瘤细胞,培养的;细胞分化;杯状细胞
Abstract： 目的 探讨转录因子Ascl2对结肠癌细胞分化的影响.方法 采用Ascl2分子干扰质粒和对照质粒对结肠癌HT -29和LS 174T细胞进行转染,通过G418筛选建立稳定转染的细胞系,采用RT-PCR和Western blot方法检测干扰Asel2分子表达对肠杯状细胞标志物Muc2和TFF3表达的影响.结果 成功构建了shRNA-Ascl2/EGFP质粒以及对照质粒shRNA-control/EGFP,经测序与预期相符,G418筛选获得shRNA-Ascl2/HT-29、shRNA-Ctr/HT-29、shRNA-Ascl2/LS174T和shRNA-Ctr/LS174T稳定转染细胞.RT-PCR和Western blot检测证实干扰质粒能够有效地抑制HT-29和LS174T细胞内的Ascl2的表达(P＜0.01).RT-PCR检测发现Muc2和TFF3 mRNA表达水平在Ascl2干扰后的HT-29细胞内均明显上调(P＜0.05)；Muc2 mRNA表达水平在Ascl2干扰后的LS174T细胞内明显上调,差异有统计学意义(P＜0.05),但是TFF3 mRNA表达水平在Ascl2干扰后的LS174T内升高不明显(P＞0.05).Western blot检测发现Muc2蛋白质表达水平在Ascl2干扰后明显上调(P＜0.05).结论 干扰结肠癌HT-29和LS174T细胞内Ascl2的表达导致其向杯状细胞的表型分化.
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