[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 Si1LI Zhong-hui2aWANG 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-198E 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