Interference of Asc12 expression in colon cancer epithelial cells leads to their differentiation into phenotype of goblet cells

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Interference of Asc12 expression in colon cancer epithelial cells leads to their differentiation into phenotype of goblet cells

更新时间：2026-01-09

- Interference of Asc12 expression in colon cancer epithelial cells leads to their differentiation into phenotype of goblet cells
- Journal of Army Medical University Vol. 34, Issue 19, Pages: 1936-1939(2012)
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  CLC： R394.2;R730.23;R735.35
- Published：2012
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李晓桓,朱蓉,田音,汪荣泉.干扰结肠癌上皮细胞Ascl2的表达致其向杯状细胞表型分化[J].陆军军医大学学报(原第三军医大学学报),2012,34(19):1936-1939.

　Li Xiaohuan,Zhu Rong,Tian Yin,Wang Rongquan.Interference of Asc12 expression in colon cancer epithelial cells leads to their differentiation into phenotype of goblet cells[J].J Amry Med Univ (J Third Mil Med Univ),2012,34(19):1936-1939.
李晓桓,朱蓉,田音,汪荣泉.干扰结肠癌上皮细胞Ascl2的表达致其向杯状细胞表型分化[J].陆军军医大学学报(原第三军医大学学报),2012,34(19):1936-1939. DOI：

　Li Xiaohuan,Zhu Rong,Tian Yin,Wang Rongquan.Interference of Asc12 expression in colon cancer epithelial cells leads to their differentiation into phenotype of goblet cells[J].J Amry Med Univ (J Third Mil Med Univ),2012,34(19):1936-1939. DOI：

摘要

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LS174T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Ascl2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（P0.01)。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（P0.05）；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（P0.05），但是TFF3 mRNA表达水平在Ascl2干扰后的LS174T内升高不明显（P0.05）。Western blot检测发现Muc2蛋白质表达水平在Ascl2干扰后明显上调（P0.05）。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。

Abstract

Objective To determine the effect of transcription factor Ascl2 on differentiation of colon cancer cells. Methods Colon cancer HT-29 and LS174T cells were transfected with shRNA-Ascl2 interference plasmid and shRNA-control plasmid

respectively. A stably-transfected cell line model was established by screening G418. Effect of shRNA-Ascl2 interference plasmid on expression of goblet cell markers

Muc2 and TFF3 mRNA and protein expression

was detected by RT-PCR and Western blot analysis. Results The successfully constructed shRNA-Ascl2/EPGF plasmid and shRNA-control/EPGF plasmid were confirmed by sequencing. Stably-transfected shRNA-Ascl2/HT-29

shRNA-Ctr/HT-29

shRNA-Ascl2/LS174T and shRNA-Ctr/LS174T cells were obtained by screening G418. RT-PCR and Western blot analysis displayed that the Ascl2 interference plasmid could effectively inhibit the Ascl2 expression in HT-29 and LS174T cells (P0.01) and the Muc2 and TFF3 mRNA expression levels were significantly higher in HT-29 and LS174T cells than in their controls after Ascl2 interference (P0.05). However

no significant difference was observed in the TFF3 mRNA expression level in LS174T cells and controls after Ascl2 interference (P0.05). Western blot analysis showed that the Muc2 protein expression level was significantly higher in shRNA-Ascl2/HT-29 and shRNA-Ascl2/LS174T cells than in their controls after Ascl2 interference (P0.05). Conclusion Interference of Ascl2 expression in colon cancer HT-29 and LS174T cells leads to their differentiation into phenotype of goblet cells.

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