Core 2 and Core 4 O-glycan chains participate in adhesion and invasion of E.coli to intestinal epithelial cells in vitro

刘韵; 叶钧; 宋丽丽; 田音; 潘琼; 彭志红; 汪荣泉

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Core 2 and Core 4 O-glycan chains participate in adhesion and invasion of E.coli to intestinal epithelial cells in vitro

更新时间：2026-01-09

- Core 2 and Core 4 O-glycan chains participate in adhesion and invasion of E.coli to intestinal epithelial cells in vitro
- Journal of Army Medical University Vol. 36, Issue 2, Pages: 105-109(2014)
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  CLC： R322.45;R363;R378.21
- Published：2014
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刘韵,叶钧,宋丽丽,田音,潘琼,彭志红,汪荣泉.Core 2和Core 4 O-型糖链参与大肠杆菌对肠上皮细胞的黏附与侵袭[J].陆军军医大学学报(原第三军医大学学报),2014,36(02):105-109.

　Liu Yun,Ye Jun,Song Lili,Tian Yin,Pan Qiong,Peng Zhihong,Wang Rongquan.Core 2 and Core 4 O-glycan chains participate in adhesion and invasion of E.coli to intestinal epithelial cells in vitro[J].J Amry Med Univ (J Third Mil Med Univ),2014,36(02):105-109.
刘韵,叶钧,宋丽丽,田音,潘琼,彭志红,汪荣泉.Core 2和Core 4 O-型糖链参与大肠杆菌对肠上皮细胞的黏附与侵袭[J].陆军军医大学学报(原第三军医大学学报),2014,36(02):105-109. DOI：

　Liu Yun,Ye Jun,Song Lili,Tian Yin,Pan Qiong,Peng Zhihong,Wang Rongquan.Core 2 and Core 4 O-glycan chains participate in adhesion and invasion of E.coli to intestinal epithelial cells in vitro[J].J Amry Med Univ (J Third Mil Med Univ),2014,36(02):105-109. DOI：

摘要

目的探讨大肠杆菌对Core 2和Core 4 O-型糖链合成障碍的肠上皮细胞的黏附和侵袭的影响。方法采用针对C2GnT-2的干扰质粒和对照质粒对结肠癌HT-29细胞进行转染，通过G418筛选建立稳定转染的细胞系，采用Real-time PCR和Western blot方法检测干扰效率，选取干扰最有效的稳定细胞系与肠致病性大肠杆菌（enteropathogenic Escherichia coli，EPEC）和肠出血性大肠杆菌（EHEC O157∶H7）37 ℃共培养。最后采用系列稀释克隆计数法观察干扰Core 2和Core 4 O-型糖链合成的C2GnT-2的HT-29细胞对细菌黏附及侵袭的影响。结果成功筛选获得shRNA-C2GnT-2及shRNA-Ctr稳定转染的HT-29细胞。Real-time PCR和Western blot检测证实干扰质粒能够有效地抑制HT-29细胞内C2GnT-2的mRNA及蛋白水平表达(P0.01)，黏附于shRNA-C2GnT-2/HT-29细胞表面的EPEC或EHEC O157∶H7的数量较对照细胞显著减少（

0.01，

0.05)，而侵袭入shRNA-C2GnT-2/HT-29细胞内的EPEC或EHEC O157∶H7的数量较对照细胞显著增加（

0.01，

0.05)。结论 Core 2和Core 4 O-型糖链参与了肠上皮细胞细菌的黏附与侵袭。

Abstract

Objective To determine the effect of mucin type Core 2 and Core 4 O-glycans on the E.coli adhesion and invasion to intestinal epithelial cells. Methods Colon cancer epithelial HT-29 cells were transfected with shRNA-C2GnT-2 interfering plasmid and shRNA-Ctr control plasmid

respectively. A stably-transfected cell line was established by G418 screening. C2GnT-2 interfering

efficiency was confirmed by real-time PCR and Western blotting. The stably-transfected cell line having best interfering effects was incubated with enteropathogenic E.coli (EPEC) or enterohemorrhagic E.coli (EHEC O157∶H7). Serial dilution cloning counting was applied to evaluate the effects of Core 2 and Core 4 O-glycan deficiency on E.coli adhesion and invasion to intestinal epithelial cells. Results The shRNA-C2GnT-2 and shRNA-Ctr stably-transfected HT-29 cells were obtained by screening with G418. Real-time PCR and Western blot results displayed that C2GnT-2 interfering could effectively inhibit C2GnT-2 expression in HT-29 cells at both mRNA and protein levels (

0.01). Adhesion of EPEC or EHEC O157∶H7 to shRNA-C2GnT-2/HT-29 cells was significantly decreased as compared to shRNA-Ctr/HT-29 or HT-29 cells (

0.01 or

0.05)

while the numbers of EPEC or EHEC O157∶H7 invaded into the shRNA-C2GnT-2/HT-29 cells were significantly higher than those into shRNA-Ctr/HT-29 or HT-29 cells (

0.01 or

0.05). Conclusion Core 2 and Core 4 O-glycan chains participate in the adhesion and invasion of EPEC or EHEC O157∶H7 to intestinal epithelial cells.

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