MiRNA-200b reverses drug resistance of hepatocellular carcinoma HepG2 cells to sorafenib and its underlying mechanism

Yong DENG; Tengqian TANG; Leida ZHANG

doi:10.16016/j.1000-5404.202105085

您当前的位置：

首页 >

文章列表页 >

MiRNA-200b reverses drug resistance of hepatocellular carcinoma HepG2 cells to sorafenib and its underlying mechanism

Basic Medicine | 更新时间：2026-01-09

- MiRNA-200b reverses drug resistance of hepatocellular carcinoma HepG2 cells to sorafenib and its underlying mechanism
- Journal of Third Military Medical University Vol. 43, Issue 22, Pages: 2435-2440(2021)
- 作者机构：
  
  400038 重庆，陆军军医大学（第三军医大学）第一附属医院肝胆外科
- 作者简介：
- 基金信息：
- DOI：10.16016/j.1000-5404.202105085
  CLC： R735.7;R915;R979.1
- Received：20 May 2021，
  
  Revised：2021-08-02，
  
  Published：30 November 2021
- 稿件说明：
移动端阅览
Yong DENG, Tengqian TANG, Leida ZHANG. MiRNA-200b reverses drug resistance of hepatocellular carcinoma HepG2 cells to sorafenib and its underlying mechanism[J]. Journal of Third Military Medical University, 2021, 43(22): 2435-2440.
DOI：

Yong DENG, Tengqian TANG, Leida ZHANG. MiRNA-200b reverses drug resistance of hepatocellular carcinoma HepG2 cells to sorafenib and its underlying mechanism[J]. Journal of Third Military Medical University, 2021, 43(22): 2435-2440. DOI： 10.16016/j.1000-5404.202105085.

摘要

目的

探究miRNA-200b对肝细胞癌索拉菲尼耐药的影响，以期延缓或改善晚期肝癌患者对索拉菲尼的耐药，增加索拉菲尼的疗效。

方法

通过逐步提高培养基中索拉菲尼浓度的方法建立索拉菲尼耐药的肝细胞癌HepG2-SR细胞系，并通过实时荧光定量（RT-PCR）法检测HepG2-SR细胞系和其母本HepG2细胞系中miRNA-200b的表达差异，同时检测HepG2-SR细胞转染miRNA-200b mimics后miRNA-200b表达的变化；CCK-8法检测转染后HepG2-SR细胞对索拉菲尼敏感度的改变；划痕实验和transwell实验检测转染前后HepG2-SR细胞迁移和侵袭能力的改变；流式细胞仪检测转染前后HepG2-SR细胞凋亡情况；Western-blot检测转染前后上皮间质转变（epithelial mesenchymal transition

EMT）标志物的改变。

结果

耐药株HepG2-SR细胞对索拉菲尼的耐药性显著高于其母本株HepG2，miRNA-200b的表达显著低于其母本株HepG2；HepG2-SR细胞转染miRNA-200b mimics后miRNA-200b表达显著升高，索拉菲尼半抑制浓度（IC

）显著降低，细胞迁移率显著降低，细胞侵袭能力显著降低，细胞凋亡显著增加，上皮细胞标志E-Cadherin表达增加，间质细胞标志N-Cadherin、Vimentin表达减弱。

结论

miRNA-200b抑制HepG2-SR细胞的增殖、迁移、侵袭和EMT，促进细胞凋亡，从而逆转索拉菲尼耐药性。

Abstract

Objective

To explore the effect of miRNA-200b on sorafenib resistance in hepatocellular carcinoma cells in order to increase the efficacy of the drug in advanced liver cancer patients.

Methods

Sorafenib resistant hepatocellular carcinoma HepG2-SR cell line was established by gradually increasing the concentration of sorafenib in the culture medium. The expression of miRNA-200b in HepG2-SR cells

HepG2 cells and miRNA-200b transfected HepG2-SR cells was detected by real-time fluorescence quantitative assay (RT-PCR). In HepG2-SR cells before and after transfection

sorafenib sensitivity was detected by CCK-8 assay

migration and invasion abilities were detected by wound-healing assay and Transwell assay

cell apoptosis was measured by flow cytometry

and expression levels of epithelial mesenchymal transition (EMT) markers were studied by Western blotting.

Results

Sorafenib resistance was significantly higher and expression of miRNA-200b was significantly lower in HepG2-SR cells than HepG2 cells. Transfection of miRNA-200b mimics resulted in significantly increased miRNA-200b expression

decreased IC

of sorafenib

reduced cell migration rate and cell invasion

enhanced apoptosis and increased expression of E-cadherin but decreased expression of N-cadherin and Vimentin.

Conclusion

MiRNA-200b reversed sorafenib resistance of HepG2-SR cells by inducing apoptosis and inhibiting cell proliferation

migration

invasion and EMT.

关键词

Keywords

references

BRAY F, FERLAY J, SOERJOMATARAM I, et al . Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries[J ] . CA Cancer J Clin, 2018, 68(6): 394-424. DOI: 10.3322/caac.21492.

BALOGH J, VICTOR D, ASHAM E H, et al . Hepatocellular carcinoma: a review[J ] . J Hepatocell Carcinoma, 2016, 3: 41-53. DOI: 10.2147/jhc.s61146.

FORNER A, REIG M, BRUIX J. Hepatocellular carcinoma[J]. Lancet, 2018, 391(10127): 1301-1314. DOI: 10.1016/s0140-6736(18)30010-2.

CHENG A L, KANG Y K, CHEN Z, et al . Efficacy and safety of sorafenib in patients in the Asia-Pacific region with advanced hepatocellular carcinoma: a phase Ⅲ randomised, double-blind, placebo-controlled trial[J ] . Lancet Oncol, 2009, 10(1): 25-34. DOI: 10.1016/s1470-2045(08)70285-7.

BRUIX J, RAOUL J L, SHERMAN M, et al . Efficacy and safety of sorafenib in patients with advanced hepatocellular carcinoma: subanalyses of a phase Ⅲ trial[J ] . J Hepatol, 2012, 57(4): 821-829. DOI: 10.1016/j.jhep.2012.06.014.

BERASAIN C. Hepatocellular carcinoma and sorafenib: too many resistance mechanisms?[J]. Gut, 2013, 62(12): 1674-1675. DOI: 10.1136/gutjnl-2013-304564.

LU T X, ROTHENBERG M E. MicroRNA[J]. J Allergy Clin Immunol, 2018, 141(4): 1202-1207. DOI: 10.1016/j.jaci.2017.08.034.

SI W, SHEN J, ZHENG H, et al . The role and mechanisms of action of microRNAs in cancer drug re sistance[J ] . Clin Epigenetics, 2019, 11(1): 25. DOI: 10.1186/s13148-018-0587-8.

GOLLAVILLI P N, PARMA B, SIDDIQUI A, et al . The role of miRNA-200b/c in balancing EMT and proliferation revealed by an activity reporter[J ] . Oncogene, 2021, 40(12): 2309-2322. DOI: 10.1038/s41388-021-01708-6.

PIPERIGKOU Z, FRANCHI M, RIETHMÜLLER C, et al . MiRNA-200b restrains EMT and aggressiveness and regulates matrix composition depending on ER status and signaling in mammary cancer[J ] . Matrix Biol Plus, 2020, 6/7: 100024. DOI: 10.1016/j.mbplus.2020.100024.

FENG B, WANG R, CHEN L B. Review of miRNA-200b and cancer chemosensitivity[J]. Biomed Pharmacother, 2012, 66(6): 397-402. DOI: 10.1016/j.biopha.2012.06.002.

POGRIBNY I P, FILKOWSKI J N, TRYNDYAK V P, et al . Alterations of microRNAs and their targets are associated with acquired resistance of MCF-7 breast cancer cells to cisplatin[J ] . Int J Cancer, 2010, 127(8): 1785-1794. DOI: 10.1002/ijc.25191.

LIU J, ZHANG X, HUANG Y, et al . MiRNA-200b and miRNA-200c co-contribute to the cisplatin sensitivity of ovarian cancer cell s by targeting DNA methyltransferases[J ] . Oncol Lett, 2019, 17(2): 1453-1460. DOI: 10.3892/ol.2018.9745.

LV Z, WEI J L, YOU W X, et al . Disruption of the c-Myc/miRNA-200b-3p/PRDX2 regulatory loop enhances tumor metastasis and chemotherapeutic resistance in colorectal cancer[J ] . J Transl Med, 2017, 15(1): 257. DOI: 10.1186/s12967-017-1357-7.

HAIDER T, PANDEY V, BANJARE N, et al . Drug resistance in cancer: mechanisms and tackling strategies[J ] . Pharmacol Rep, 2020, 72(5): 1125-1151. DOI: 10.1007/s43440-020-00138-7.

GIOVANNETTI E, EROZENCI A, SMIT J, et al . Molecular mechanisms underlying the role of microRNAs (miRNAs) in anticancer drug resistance and implications for clinical practice[J ] . Crit Rev Oncol Hematol, 2012, 81(2): 103-122. DOI: 10.1016/j.critrevonc.2011.03.010.

CHEN L, WANG X, ZHU Y, et al . miR-200b-3p inhibits proliferation and induces apoptosis in colorectal cancer by targeting Wnt1[J ] . Mol Med Rep, 2018, 18(3): 2571-2580. DOI: 10.3892/mmr.2018.9287.

WANG B G, LI J S, LIU Y F, et al . MicroRNA-200b suppresses the invasion and migration of hepatocellular carcinoma by downregulating RhoA and circRNA_000839[J ] . Tumour Biol, 2017, 39(7): 1010428317719577. DOI: 10.1177/1010428317719577.

THIERY J P, ACLOQUE H, HUANG R Y, et al . Epithelial-mesenchymal transitions in development and disease[J ] . Cell, 2009, 139(5): 871-890. DOI: 10.1016/j.cell.2009.11.007.

HUNG C S, LIU H H, LIU J J, et al . MicroRNA-200a and -200b mediated hepatocellular carcinoma cell migration through the epithelial to mesenchymal transition markers[J ] . Ann Surg Oncol, 2013, 20(3): 360-368. DOI: 10.1245/s10434-012-2482-4.

TSAI S C, LIN C C, SHIH T C, et al . The miRNA-200b-ZEB1 circuit regulates diverse stemness of human hepatocellular carcinoma[J ] . Mol Carcinog, 2017, 56(9): 2035-2047. DOI: 10.1002/mc.22657.

Views

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

Sorafenib induces aggregation of macrophages and natural killer cells in hepatocellular carcinoma rats

SLC38A1 drives malignant progression of hepatocellular carcinoma by activating PI3K/AKT signaling via enhanced glutamine uptake

Emerging advances in clinical diagnosis and treatment of hepatocellular carcinoma: evolution of comprehensive treatment strategies

PD-L1 promotes hepatocellular carcinoma progression by inhibiting PTP1B and activating FAK

Nuclear EGFR cooperates with YY1 to drive CEMIP expression and enhance hepatocellular carcinoma invasion

Related Author

许文

王阁

陈川

张志敏

李琼

雒喜忠

郑继军

胡庆

Related Institution

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

Digestive Disease Center， Chongqing Hygeia Hospital

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

No.953，Xigaze， Tibet Autonomous Region

Department of Biochemistry and Molecular Biology

⁰