[1]李晓桓,朱蓉,田音,等.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,et al.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.
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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(/HTML )
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陆军军医大学学报(原第三军医大学学报)[ISSN:1000-5404/CN:51-1095/R]

卷:
34卷
期数:
2012年第19期
页码:
1936-1939
栏目:
论著
出版日期:
2012-10-15

文章信息/Info

Title:
Interference of Asc12 expression in colon cancer epithelial cells leads to their differentiation into phenotype of goblet cells
作者:
李晓桓朱蓉田音汪荣泉
第三军医大学西南医院全军消化病研究所
Author(s):
Li Xiaohuan Zhu Rong Tian Yin Wang Rongquan
Institute of Gastroenterology, Southwest Hospital, Third Military Medical University, Chongqing, 400038, China
关键词:
Ascl2结肠肿瘤肿瘤细胞 培养的细胞分化杯状细胞
Keywords:
Ascl2colon tumortumor cells cultured cell differentiation goblet cells
分类号:
R394.2; R730.23; R735.35
文献标志码:
A
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。
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 (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-Ascl2/HT-29 and shRNA-Ascl2/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.

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更新日期/Last Update: 2012-09-27