STAT3通路参与白细胞介素-22促进人结肠癌细胞系HT29紧密连接蛋白的表达

基础医学与临床 ›› 2021, Vol. 41 ›› Issue (6): 792-798.

STAT3通路参与白细胞介素-22促进人结肠癌细胞系HT29紧密连接蛋白的表达

宋茜¹, 黄雪^1*, 覃蒙斌², 张君红¹, 井洁¹, 罗毅华¹, 李雨芯¹

1.广西医科大学第一附属医院老年病学消化科, 广西壮族自治区南宁 530021;
2.广西医科大学第二附属医院消化内科,广西壮族自治区南宁 530000

收稿日期:2021-01-07 修回日期:2021-04-16 出版日期:2021-06-05 发布日期:2021-05-31
通讯作者: ^*hb960305@163.com
基金资助:
国家自然科学基金(81660093);广西研究生教育创新计划(YCSW2020112)

STAT3 pathway is involved in interleukin-22 promoting expression of tight junction protein in human colorectal cancer cell line HT29

SONG Qian¹, HUANG Xue^1*, QIN Meng-bin², ZHANG Jun-hong¹, JING Jie¹, LUO Yi-hua¹, LI Yu-xin¹

1. Department of Geriatric Gastroenterology, the First Affiliated Hospital, Guangxi Medical University, Nanning 530021;
2. Department of Gastroenterology, the Second Affiliated Hospital, Guangxi Medical University,Nanning 530000, China

Received:2021-01-07 Revised:2021-04-16 Online:2021-06-05 Published:2021-05-31
Contact: ^*hb960305@163.com

摘要/Abstract

摘要： 目的探讨白细胞介素-22(IL-22)对肠上皮屏障紧密连接的影响及机制。方法将人结肠癌细胞系HT29分为对照组、IL-22组(100 ng/mL)、FLLL32组(IL-22干预前使用FLLL32预处理)、colivelin组(IL-22干预前使用colivelin预处理)。转染处理组:shNC组、shNC+ IL-22组(IL-22干预前转染阴性对照)、shSTAT3组(转染LV-STAT3-RNAi)、shSTAT3+ IL-22组(IL-22干预前转染LV-STAT3-RNAi)。用Western blot、real-time PCR检测HT29细胞 STAT3、p-STAT3和紧密连接蛋白ZO-1、occludin、claudin-1和claudin-2及其mRNA的表达;用透射电镜观察HT29细胞紧密连接结构。结果相比于对照组,IL-22组的STAT3、p-STAT3、 ZO-1、occludin、claudin-1和claudin-2蛋白及mRNA表达水平明显升高(P＜0.05),且紧密连接更致密。相比于IL-22组,FLLL32组中上述指标的蛋白及mRNA表达量明显下调(P＜0.05),紧密连接较差,colivelin组则表达上调(P＜0.05),紧密连接完好。相比于shNC组,shNC+ IL-22组的所有紧密连接的蛋白及mRNA表达量明显增加(P＜0.05),紧密连接结构更致密;而相比于shSTAT3组,shSTAT3+ IL-22组的所有紧密连接的蛋白及mRNA表达量的差异无统计学意义。结论 STAT3通路参与IL-22促进HT29细胞紧密连接蛋白ZO-1、occludin、claudin-1和claudin-2的表达。

关键词: 紧密连接蛋白, 白细胞介素-22, STAT3, 溃疡性结肠炎, 肠上皮屏障

Abstract: Objective To investigate the effect of interleukin-22(IL-22) on intestinal epithelial barrier tight junction and its mechanism. Methods Human colon cancer HT29 cells were divided into control group, IL-22 group(100 ng/mL), FLLL32 group(pretreatment with FLLL32 before IL-22 intervention) and colivelin group(pretreatment with colivelin before IL-22 intervention). The transfection treatment groups included: shNC group, shNC+ IL-22 group(infection with negative control virus before IL-22 intervention), shSTAT3 group(infection with LV-STAT3-RNAi virus), and shSTAT3+ IL-22 group(LV-STAT3-RNAi virus infection before IL-22 intervention). Western blot and real-time PCR were used to detect the protein and mRNA expression of STAT3, p-STAT3 tight junction protein ZO-1, occludin, claudin-1 and claudin-2 in HT29 cells. Transmission electron microscopy(TEM) was used to observe the changes in the structure of tight junctions of HT29 cells. Results The protein and mRNA expression level of STAT3, p-STAT3, ZO-1, occludin, claudin-1 and claudin-2 in IL-22 group were significantly higher than those in the control group(P＜0.05) with more compact, tight junction. Compared with IL-22 group, protein and mRNA expression level of the above indexes in FLLL32 group were significantly decreased(P＜0.05), the tight junction was poor, the expression of colivelin group was up-regulated(P＜0.05), the tight junction was intact. The shNC+ IL-22 group significantly increased the expression of all tightly bound proteins and mRNA compared with the shNC group(P＜0.05), the tight junction structure was more dense, while the shSTAT3+ IL-22 group showed no significant difference in protein and mRNA expression level when compared with theose of shSTAT3 group. Conclusions STAT3 signaling pathway is involved in IL-22 to promote the expression of tight junction proteins ZO-1, occludin, claudin-1 and claudin-2 in HT29 cells.

Key words: tight junction protein, IL-22, STAT3, ulcerative colitis, intestinal epithelial barrier

中图分类号:

R319

宋茜, 黄雪, 覃蒙斌, 张君红, 井洁, 罗毅华, 李雨芯. STAT3通路参与白细胞介素-22促进人结肠癌细胞系HT29紧密连接蛋白的表达[J]. 基础医学与临床, 2021, 41(6): 792-798.

SONG Qian, HUANG Xue, QIN Meng-bin, ZHANG Jun-hong, JING Jie, LUO Yi-hua, LI Yu-xin. STAT3 pathway is involved in interleukin-22 promoting expression of tight junction protein in human colorectal cancer cell line HT29[J]. Basic & Clinical Medicine, 2021, 41(6): 792-798.

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