[1]刘韵,叶钧,宋丽丽,等.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].第三军医大学学报,2014,36(02):105-109.
 Liu Yun,Ye Jun,Song Lili,et al.Core 2 and Core 4 O-glycan chains participate in adhesion and invasion of E.coli to intestinal epithelial cells in vitro[J].J Third Mil Med Univ,2014,36(02):105-109.
点击复制

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(/HTML )
分享到:

《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
36卷
期数:
2014年第02期
页码:
105-109
栏目:
论著
出版日期:
2014-01-30

文章信息/Info

Title:
Core 2 and Core 4 O-glycan chains participate in adhesion and invasion of E.coli to intestinal epithelial cells in vitro
作者:
刘韵叶钧宋丽丽田音潘琼彭志红汪荣泉
第三军医大学西南医院全军消化病研究所
Author(s):
Liu Yun Ye Jun Song Lili Tian Yin Pan Qiong Peng Zhihong Wang Rongquan
Institute of Gastroenterology, Southwest Hospital, Third Military Medical University, Chongqing, 400038, China
关键词:
黏蛋白大肠杆菌O-型糖链C2GnT-2细胞黏附侵袭
Keywords:
mucin E.coli O-glycan C2GnT-2 cell adhesion invasion
分类号:
R322.45;R363;R378.21
文献标志码:
A
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(P<0.01),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(P<0.01,P<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(P<0.01,P<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 (P<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 (P<0.01 or P<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 (P<0.01 or P<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.

相似文献/References:

[1]陈曦,黄悦,于彬,等.结直肠癌单克隆抗体的制备及生物学特性鉴定[J].第三军医大学学报,2007,29(10):896.
 CHEN Xi,HUANG Yue,YU Bin,et al.Preparing and identifying a monoclonal antibody against human colorectal carcinoma[J].J Third Mil Med Univ,2007,29(02):896.
[2]胡沛臻,温江田,路凡,等.溶氧反馈-补料分批技术高密度培养基因重组MAGE1/HSP70/MAGE3工程菌[J].第三军医大学学报,2007,29(16):1576.
 HU Pei-zhen,WEN Jiang-tian,LU Fan,et al.Preparation of gene recombinant MAGE1/HSP70/MAGE3 by high cell density culture of E.coli. with DO fed-back control of nutrient feeding[J].J Third Mil Med Univ,2007,29(02):1576.
[3]刘建,程浩,邹姝丽,等.宫内给药对大鼠细菌性宫内感染胎盘IL-6与MMP-9表达的影响[J].第三军医大学学报,2008,30(19):1803.
 LIU Jian,CHENG Hao,ZOU Shu-li,et al.Intraamniotic administration on placental IL-6 and MMP-9 expressions in rats with intrauterine infection of E.coli[J].J Third Mil Med Univ,2008,30(02):1803.
[4]邹锋,郝飞,宋志强,等.HSPC016基因重组工程菌发酵研究[J].第三军医大学学报,2005,27(09):904.
[5]孙凤军,枉前,史惠卿,等.luxS基因敲除临床分离大肠杆菌株生物膜形成能力的变化[J].第三军医大学学报,2011,33(03):258.
 Sun Fengjun,Wang Qian,Shi Huiqing,et al.Effect of luxS gene on biofilm formation in knocked-out clinically isolated Escherichia coli[J].J Third Mil Med Univ,2011,33(02):258.
[6]何莉,鲜尽红,刘高科,等.重组工程菌pET32a-CR1-SCR15-18/BL21(DE3)高密度发酵工艺研究[J].第三军医大学学报,2011,33(09):896.
 He Li,Xian Jinhong,Liu Gaoke,et al.High density fermentation of recombinant bacterium pET32a-CR1-SCR15-18/BL21(DE3)[J].J Third Mil Med Univ,2011,33(02):896.
[7]陈盛,余加林,罗则佳,等.巯乙磺酸钠单独及联合环丙沙星抗大肠杆菌生物膜的作用[J].第三军医大学学报,2012,34(06):488.
 Chen Sheng,Yu Jialin,Luo Zejia,et al.Effects of mesna alone or combined with ciprofloxacin on Escherichia coli biofilm[J].J Third Mil Med Univ,2012,34(02):488.
[8]陈盛,余加林,罗则佳,等.巯乙磺酸钠对家兔留置导尿管表面大肠杆菌生物膜的作用[J].第三军医大学学报,2012,34(10):921.
 Chen Sheng,Yu Jialin,Luo Zejia,et al.Effect of mesna on Escherichia coli biofilm on indwelling urethral catheter in rabbits[J].J Third Mil Med Univ,2012,34(02):921.
[9]陈盛,余加林,何念海,等.巯乙磺酸钠对大肠杆菌生物膜早期黏附及胞外聚合物的影响[J].第三军医大学学报,2011,33(24):2554.
 Chen Sheng,Yu Jialin,He Nianhai,et al.Effects of mesna on Escherichia coli biofilm adhesion and extracellular polymeric substances in vitro[J].J Third Mil Med Univ,2011,33(02):2554.
[10]刘正祥,邹全明,洪愉,等.重组幽门螺杆菌粘附素工程菌高密度发酵条件的研究[J].第三军医大学学报,2004,26(15):0.[doi:10.16016/j.1000-5404.2004.15.018 ]
 LIU Zheng xiang,ZOU Quan ming,HONG Yu,et al.[J].J Third Mil Med Univ,2004,26(02):0.[doi:10.16016/j.1000-5404.2004.15.018 ]

更新日期/Last Update: 2014-01-21