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干扰结肠癌上皮细胞Ascl2的表达致其向杯状细胞表型分化 被引量：6

Interference of Asc12 expression in colon cancer epithelial cells leads to their differen-tiation into phenotype of goblet cells
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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(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。 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 interfer- ence plasmid and shRNA-control plasmid, respectively. A stably-transfected cell line model was established by screening G418. Effect of shRNA-Ascl2 interfe-rence 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-Asc12/HT-29, shRNA-Ctr/HT-29, shRNA-Asc12/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 （P 〈 0.01 ） and the Muc2 and TFF3 mRNA expression levels were significantly higher in HT-29 and LS174T cells than in their controls after Ascl2 interference （P 〈 0.05 ）. However, no significant difference was observed in the TFF3 mRNA expression level in LS174T cells and controls after Ascl2 interference （P 〉 0.05）. Western blot analysis showed that the Muc2 protein expression level was significantly higher in shRNA-Asc12/HT-29 and shRNA-Asc12/LS174T cells than in their controls after Ascl2 interference （P 〈 0.05 ）. Conclusion Interference of Ascl2 expression in colon cancer HT-29 and LS174T cells leads to their differentiation into phenotype of goblet cells.
作者 李晓桓 朱蓉 田音 汪荣泉
机构地区 第三军医大学西南医院全军消化病研究所
出处 《第三军医大学学报》 CAS CSCD 北大核心 2012年第19期1936-1939,共4页 Journal of Third Military Medical University
关键词 Ascl2 结肠肿瘤 肿瘤细胞 培养的 细胞分化 杯状细胞 Ascl2 colon tumor tumor cells, cultured cell differentiation goblet cellsCorresponding author ： Wang Rongquan, Tel ： 86 -23-68754124, E-mail ： rongquanw@ hotmail, com
分类号 R394.2 [医药卫生—医学遗传学] R730.23 [医药卫生—肿瘤]
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参考文献14

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同被引文献84

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第三军医大学学报
2012年 第19期

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