[1]陈 偲,李忠辉a,王 颖b.miR-198 通过靶向ZEB2 调控EMT 过程抑制肝癌细胞增殖和迁移的机制研究[J].现代检验医学杂志,2022,37(04):23-29.[doi:10.3969/j.issn.1671-7414.2022.04.005]
　CHEN Si,LI Zhong-huia,WANG Yingb.Study on the Mechanism of miR-198 Inhibiting the Proliferation and Migration of Hepatoma Cells by Regulating EMT Process by Targeting ZEB2[J].Journal of Modern Laboratory Medicine,2022,37(04):23-29.[doi:10.3969/j.issn.1671-7414.2022.04.005]
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miR-198 通过靶向ZEB2 调控EMT 过程抑制肝癌细胞增殖和迁移的机制研究()
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《现代检验医学杂志》[ISSN:/CN:]

卷:
第37卷
期数:
2022年04期
页码:
23-29
栏目:
论著
出版日期:
2022-07-15

文章信息/Info

Title:
Study on the Mechanism of miR-198 Inhibiting the Proliferation and Migration of Hepatoma Cells by Regulating EMT Process by Targeting ZEB2
文章编号:
1671-7414（2022）04-023-07
作者:
陈 偲1; 李忠辉2a; 王 颖2b
1. 解放军联勤保障部队第九六九医院感染科, 呼和浩特 010010；2. 内蒙古包头市昆都仑区包钢医院a. 肿瘤内科；b. 老年病科，内蒙古包头 014010
Author(s):
CHEN Si1; LI Zhong-hui2a; WANG Ying2b
1. Department of Infectious Disease, PLA Joint Support Unit 969 Hospital, Hohhot 010010，China；2a. Department of Oncology；2b. Department of Geriatric, Inner Mongolia Baotou Kundulun District Baogang Hospital, Inner Mongolia Baotou 010010，China
关键词:
肝细胞癌; 微小核糖核酸-198; E 盒结合锌指蛋白2 (ZEB2); 增殖; 迁移; 上皮间质转化
分类号:
R735.7；R730.43
DOI:
10.3969/j.issn.1671-7414.2022.04.005
文献标志码:
A
摘要:
目的 探讨微小核糖核酸-198（miR-198）在肝细胞癌（hepatocellular carcinoma，HCC）组织细胞中的表达及其对癌细胞增殖、迁移的影响及其相关机制。方法 采用qRT-PCR 检测miR-198 在HCC 组织及细胞(HepG2，Hep3G，MHCC97H，Huh7) 中的表达情况；采用脂质体2000 向Huh7 细胞中单独转染miR-198 mimics，共转染miR-198 mimics 和pcDNA3.1-ZEB2；生物信息学网站预测miR-198 的潜在靶基因，采用双荧光素酶报告基因实验进行验证；采用Western blot 检测E 盒结合锌指蛋白2（E-box binding zinc finger protein 2, ZEB2）及上皮-间质转化（epithelialmesenchymaltransformation, EMT）相关蛋白表达；CCK-8 法和细胞划痕实验检测Huh7 细胞增殖和迁移能力。结果 20 例HCC 组织中miR-198相对表达量（0.354±0.022）明显低于邻近正常组织（4.762±1.135），差异有统计学意义（t=17.365，P＜ 0.001）。HCC细胞系HepG2（0.589±0.103），Hep3G（0.495±0.086），MHCC97H（0.558±0.056）和Huh7（0.362±0.045）中miR-198 相对表达量较正常肝细胞LO2（1.823±0.125）显著降低（t=17.159，18.466，17.590，20.315，均P ＜ 0.05）。miR-198 mimics 组细胞增殖（0.398±0.146 vs 0.691±0.213）和迁移能力（20.012±2.103 vs 84.032±6.512）较对照组明显降低（t=1.965，52.459，均P ＜ 0.001）。miR-198 mimics 组细胞中E-cadherin 表达较对照组明显升高，N-cadherin和Vimentin 表达较对照组明显降低（t=18.478，17.550，19.706，均P ＜ 0.01）。ZEB2 是miR-198 的靶基因，miR-198负调控ZEB2。20 例HCC 组织中ZEB2 相对表达量（3.621±1.143）明显高于邻近正常组织（ 0.736±0.030），差异有统计学意义（t=11.284，P ＜ 0.001），与miR-198 表达呈负相关（r=-0.702，P ＜ 0.05）。共转染过表达ZEB2 逆转了miR-198 mimics 对HCC 细胞增殖、迁移和EMT 的抑制作用。结论 miR-198 在HCC 组织和细胞中表达下调，miR-198 通过靶向负调控ZEB2 的表达抑制Huh7 细胞的增殖和迁移。
Abstract:
Objective To investigate the expression of micro RNA（miR）-198 in hepatocellular carcinoma (HCC) tissues and its effects on the proliferation and migration of cancer cells and related mechanisms. Methods qRT-PCR was used to detect the expression of miR-198 in HCC tissues and cells (HepG2, Hep3G, MHCC97H, Huh7). The liposome 2000 was used to transfect miR-198 mimics alone，co-transfect miR-198 mimics and pcDNA3.1-ZEB2 into Huh7 cells. Bioinformatics website predicted that the potential target gene of miR-198，and the double luciferase reporter gene experiment was used for verification. Western blot was used to detect the expression of E-box binding zinc finger protein 2（ZEB2） protein and epithelial-mesenchymal transformation (EMT) related proteins,and CCK-8 and scratch test were used to detect the proliferation and migration ability of Huh7 cells. Results The relative expression of miR-198 in 20 HCC tissues (0.354±0.022）was significantly lower than that in adjacent normal tissues（4.762±1.135), with the difference was statistically significant (t=17.365, P ＜ 0.001). HCC cell lines HepG2（0.589±0.103）, Hep3G（0.495±0.086）, MHCC97H（0.558±0.056） and Huh7（0.362±0.045） was significantly lower than that in normal liver cells LO2 miR-198 in normal liver cell（1.823±0.125）（t=17.159, 18.466, 17.590, 20.315, all P ＜ 0.05）. The proliferation(0.398±0.146 vs 0.691±0.213) and migration(20.012±2.103 vs 84.032±6.512) of miR-198 mimics group were significantly decreased compared with the control group (t=1.965, 52.459, all P ＜ 0.001). The expression of E-cadherin(2.010±0.089 vs 1.001±0.032) in miR-198 mimics group was significantly higher than that in control group, and the expression of N-cadherin(0.362±0.056 vs 1.001±0.029) and Vimentin(0.356±0.048 vs 1.000±0.030) was significantly lower than that in control group (t=18.478, 17.550, 19.706, all P ＜ 0.01). ZEB2 was the target gene of miR-198 and miR-198 negatively regulates ZEB2. The relative expression of ZEB2 in 20 HCC tissues (3.621±1.143)was significantly higher than that in adjacent normal tissues（0.736±0.030) ，the difference was statistically significant (t=11.284, P ＜ 0.001), and was negatively correlated with miR-198 expression (r= -0.702, P ＜ 0.05). Overexpression of ZEB2 by cotransfection reversed the inhibitory effect of miR-198 mimics on HCC cell proliferation, migration and EMT. Conclusion MiR-198 was down-regulated in HCC tissues and cells, and inhibits the proliferation and migration of Huh7 cells by negatively regulating the expression of ZEB2.

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备注/Memo

备注/Memo:
基金项目：内蒙古自然科学技术基金项目（2019BS08056）。
作者简介： 陈偲(1984-)，女，硕士研究生，副主任医师，研究方向：肝脏疾病的诊治，E-mail：chensipla@163.com。
通讯作者： 王颖(1978-)，女，研究生，副主任医师，研究方向：老年人恶性肿瘤的治疗，E-mail：595400820@qq.com。
更新日期/Last Update: 1900-01-01