miR-576-5p regulates the EMT process to inhibit migration and invasion of hepatocellular carcinoma cells

Qi CUI; Yuyan WANG; Yingjun LU; Aizhong ZENG

doi:10.16016/j.2097-0927.202205009

您当前的位置：

首页 >

文章列表页 >

miR-576-5p regulates the EMT process to inhibit migration and invasion of hepatocellular carcinoma cells

Basic Medicine | 更新时间：2026-01-09

- miR-576-5p regulates the EMT process to inhibit migration and invasion of hepatocellular carcinoma cells
- Journal of Army Medical University Vol. 45, Issue 4, Pages: 366-373(2023)
- 作者机构：
  
  400016 重庆，重庆医科大学附属第一医院感染科
- 作者简介：
- 基金信息：
- DOI：10.16016/j.2097-0927.202205009
  CLC： R394.3;R73-37;R735.7
- Received：04 May 2022，
  
  Revised：2022-10-26，
  
  Published：28 February 2023
- 稿件说明：
移动端阅览
Qi CUI, Yuyan WANG, Yingjun LU, et al. miR-576-5p regulates the EMT process to inhibit migration and invasion of hepatocellular carcinoma cells[J]. Journal of Army Medical University, 2023, 45(4): 366-373.
DOI：

Qi CUI, Yuyan WANG, Yingjun LU, et al. miR-576-5p regulates the EMT process to inhibit migration and invasion of hepatocellular carcinoma cells[J]. Journal of Army Medical University, 2023, 45(4): 366-373. DOI： 10.16016/j.2097-0927.202205009.

摘要

目的

探究miR-576-5p调控上皮细胞-间充质转化(epithelial-mesenchymal transition

EMT)进程在肝癌细胞迁移和侵袭中的影响。

方法

采用RT-qPCR检测重庆医科大学附属第一医院肝胆外科2020年6月至2021年1月行肝癌切除术的27例患者的肝细胞癌组织(hepatocellular carcinoma

HCC)和癌旁组织，同时检测了肝癌细胞株和正常人肝细胞的miR-576-5p表达水平。将miR-576-5p类似物/阴性对照(miR-576-5p mimics/ mimics NC)转染到hepG2作为过表达组(mimics组)和过表达对照组(mimics NC组)；将miR-576-5p抑制剂/阴性对照(miR-576-5p inhibitor/ inhibitor NC)转染至SMMC-7721中作为去表达组(inhibitor组)及去表达对照组(inhibitor NC组)。用CCK-8和EDU检测细胞增殖能力；划痕实验、Transwell侵袭实验测定各组细胞侵袭和迁移能力；Western blot检测各组细胞中E-cadherin、N-cadherin和snail蛋白表达水平变化。

结果

miR-576-5p在肝细胞癌组织中较配对癌旁组织表达明显下降(

0.001)，在肝癌细胞株中较正常人肝细胞表达偏低(

0.001)。SMMC-7721 inhibitor组较inhibitor NC组增殖能力增强(

0.001，

0.001)，伤口愈合率显著增强(

0.001)，transwell小室迁移和侵袭穿孔细胞数明显增加(

0.001，

0.01)。mimics组较过表达对照组增殖能力减弱(

0.01，

0.01)，伤口愈合率显著降低(

0.01)，迁移和侵袭穿孔细胞数明显减少(

0.001，

0.01)。miR-576-5p过表达组与其对照组比较，E-cadherin蛋白表达显著增加，N-cadherin和snail蛋白表达明显减少(

0.001，

0.05，

0.05)，miR-576-5p去表达则发生相反的变化(

0.05，

0.001

0.01)。结论miR-576-5p抑制肝癌细胞的增殖，同时阻碍EMT进程，抑制肝癌细胞侵袭和迁移。

Abstract

Objective

To investigate the effect of miR-576-5P on the migration and invasion of hepatocellular carcinoma(HCC)cells by regulating epithelial-mesenchymal transition (EMT) process.

Methods

RT-qPCR was used to detect miR-576-5p expression level in HCC cells and normal liver cells

hepatocellular carcinoma tissues and paraneoplastic tissues from 27 patients who accepted hepatectomy from June 2020 to January 2021 in the Department of Hepatobiliary Surgery of the First Affiliated Hospital of Chongqing Medical University. MiR-576-5p mimics and mimics NC were transfected into hepG2 cells as the over-expression group(mimics group) and the control group(mimics NC group)

and miR-576-5p inhibitor and inhibitor NC were transfected into SMMC-7721 cells as the de-expression group(inhibitor group) and the control group(inhibitor NC group). CCK-8 assay and EDU assay were used to detect the proliferative ability of cells. Transwell assay and wound healing assay were used to observe the changes of invasion and migration of cells in each group. The changes of expression level of E-cadherin

N-cadherin and snail in each group were detected by Western blotting.

Results

The expression of miR-576-5p in HCC tissues was significantly lower than that in matched adjacent tissues(

0.001)

and the expression of miR-576-5p was lower in HCC cells than that of normal liver cells(

0.001). SMMC-7721 inhibitor group had enhanced proliferative capacity (

0.001

0.001)

significantly increased wound healing rate (

0.001) and significantly increased migration and invasive perforated cells (

0.001

0.01)

compared to the inhibited NC group. HepG2 mimics group had lower proliferative capacity(

0.01

0.01)

significantly lower woun

d healing rate(

0.01)

and significantly fewer migration and invasive perforated cells than the mimics NC group(

0.001

0.01). The hepG2 mimics group showed a obviously increase in the expression of E-cadherin and a decrease in the expression of N-cadherin and snail compared to the mimics NC group

0.001

0.05

0.05)

while the completely opposite changes occurred in the SMMC-7721 inhibitor groups (

0.05

0.001

0.01).

Conclusion

MiR-576-5p inhibits the proliferation of hepatocellular carcinoma cells and prevents the EMT process to inhibit invasion and migration of cells.

关键词

Keywords

references

SINGAL A G, LAMPERTICO P, NAHON P. Epidemiology and surveillance for hepatocellular carcinoma: new trends[J]. J Hepatol, 2020, 72(2): 250-261. DOI: 10.1016/j.jhep.2019.08.025.

FUJIWARA N, FRIEDMAN S L, GOOSSENS N, et al . Risk factors and prevention of hepatocellular carcinoma in the era of precision medicine[J ] . J Hepatol, 2018, 68(3): 526-549. DOI: 10.1016/j.jhep.2017.09.016.

BERTINO G, NERI S, BRUNO C M, et al . Diagnostic and prognostic value of alpha-fetoprotein, des-γ-carboxy prothrombin and squamous cell carcinoma antigen immunoglobulin M complexes in hepatocellular carcinoma[J ] . Minerva Med, 2011, 102(5): 363-371.

VALINEZHAD ORANG A, SAFARALIZADEH R, KAZEMZADEH-BAVILI M. Mechanisms of miRNA-mediated gene regulation from common downregulation to mRNA-specific upregulation[J]. Int J Genomics, 2014, 2014: 970607. DOI: 10.1155/2014/970607.

VASUDEVAN S. Posttranscriptional upregulation by mRNAs[J]. Wiley Interdiscip Rev RNA, 2012, 3(3): 311-330. DOI: 10.1002/wrna.121.

HUANG V, PLACE R F, PORTNOY V, et al . Upregulation of Cyclin B1 by miRNA and its implications in cancer[J ] . Nucleic Acids Res, 2012, 40(4): 1695-1707. DOI: 10.1093/nar/gkr934.

HUANG J Y, ZHANG K, CHEN D Q, et al . microRNA-451: epithelial-mesenchymal transition inhibitor and prognostic biomarker of hepatocelluar carcinoma[J ] . Oncotarget, 2015, 6(21): 18613-18630. DOI: 10.18632/oncotarget.4317.

NIETO M A. Epithelial plasticity: a common theme in embryonic and cancercells[J]. Science, 2013, 342(6159): 1234850. DOI: 10.1126/science.1234850.

GIANNELLI G, KOUDELKOVA P, DITURI F, et al . Role of epithelial tomesenchymal transition in hepatocellular carcinoma[J ] . J Hepatol, 2016, 65(4): 798-808. DOI: 10.1016/j.jhep.2016.05.007.

ÖSTERREICHER C H, PENZ-ÖSTERREICHER M, GRI-VENNIKOV S I, et al . Fibroblast-specific protein 1 identifies an inflammatory subpopulation of macrophages in theliver[J ] . PNAS, 2011, 108(1): 308-313. DOI: 10.1073/pnas.1017547108.

LUO J L, LIU L Y, SHEN J W, et al . miR-576-5p promotes epithelial-to-mesenchymal transition in colorectal cancer by targeting the Wnt5a-mediated Wnt/β-catenin signaling pathway[J ] . Mol Med Rep, 2021, 23(2): 94. DOI: 10.3892/mmr.2020.11733.

ZHANG L Y, PAN K, ZUO Z K, et al . LINC01133 hampers the development of gastric cancer through increasing somatostatin via binding to mRNA-576-5p[J ] . Epigenomics, 2021, 13(15): 1205-1219. DOI: 10.2217/epi-2020-0377..

NI X F, ZHAO L H, LI G, et al . microRNA-548-3p and microRNA-576-5p enhance the migration and invasion of esophageal squamous cell carcinoma cells via NRIP1 down-regulation[J ] . Neoplasma, 2018, 65(6): 881-887. DOI: 10.4149/neo_2018_171206N803.

ROBERTS A P, LEWIS A P, JOPLING C L. miR-122 activates hepatitis C virus translation by a specialized mechanism requiring particular RNA components [J]. Nucleic acids research, 2011, 39(17): 7716-7729. DOI: 10.1093/nar/gkr426.

GRAMANTIERI L, FORNARI F, FERRACIN M, et al . microRNA-221 targets bmf in hepatocellular carcinoma and correlates with tumor multifocality[J ] . Clin Cancer Res, 2009, 15(16): 5073-5081. DOI: 10.1158/1078-0432.CCR-09-0092.

SUN X G, HOU Z Y, LI N, et al . miR-597-5p suppresses the progression of hepatocellular carcinoma via targeting transcriptional enhancer associate domain transcription factor 1(TEAD1)[J ] . In Vitro Cell Dev Biol Anim, 2022, 58(2): 96-108. DOI: 10.1007/s11626-021-00614-1..

PONNUSAMY M P, SESHACHARYULU P, LAKSHMANAN I, et al . Emerging role of mucins in epithelial to mesenchymal transition[J ] . Curr Cancer Drug Targets, 2013, 13(9): 945-956. DOI: 10.2174/15680096113136660100.

YUAN J H, YANG F, WANG F, et al . A long noncoding RNA activated by TGF-β promotes th e invasion-metastasis cascade in hepatocellular carcinoma[J ] . Cancer Cell, 2014, 25(5): 666-681. DOI: 10.1016/j.ccr.2014.03.010.

KIM T, VERONESE A, PICHIORRI F, et al . P53 regulates epithelial-mesenchymal transition through microRNAs targeting ZEB1 and ZEB2[J ] . J Exp Med, 2011, 208(5): 875-883. DOI: 10.1084/jem.20110235.

ZHANG J P, ZENG C, XU L, et al . MicroRNA-148a suppresses the epithelial-mesenchymal transition and metastasis of hepatoma cells by targeting Met/Snail signaling[J ] . Oncogene, 2014, 33(31): 4069-4076. DOI: 10.1038/onc.2013.369.

Views

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

ETS1 transcription up-regulates FBXO45 and promotes invasion and migration of hepatocellular carcinoma via epithelial-mesenchymal transition

Bilirubin impairs the sorafenib sensitivity in Huh7 cells through fatty acid oxidation by up-regulating PPARα/CPT1A

Mechanism of miR-409-3p negatively regulating epithelial mesenchymal transition of ovarian serous carcinoma HO-8910 cells by targeting ZEB1

HBx promotes liver tumor in mice by inhibiting microRNA-145a-5p

Hepatitis B virus X promotes tumorigenesis of hepatocellular carcinoma through HIPPO signaling pathway in mice

Related Author

ZHU Zhenbao

WU Feifan

GU Yongpeng

XIE Chuanming

ZHANG Leida

Jun TAN

Chi ZHANG

Yan DONG

Related Institution

Department of Hepatobiliary Surgery， the Second Affiliated Hospital of Chongqing Medical University

Department of Hepatobiliary Surgery， First Affiliated Hospital， Army Medical University （Third Military Medical University）

Department of Oncology, First Affiliated Hospital, Army Medical University (Third Military Medical University)

Department of Gynecology and Obstetrics, Second Affiliated Hospital, Army Medical University (Third Military Medical University)

Molecular Medicine and Cancer Research Center, Biochemistry and Molecular Biology Research Center, College of Pharmacy, Chongqing Medical University

⁰