Activation of miR-101-3p/EZH2 pathway enhances sorafenib sensitivity of HepG2 cells

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Activation of miR-101-3p/EZH2 pathway enhances sorafenib sensitivity of HepG2 cells

Basic Medicine | 更新时间：2026-01-09

- Activation of miR-101-3p/EZH2 pathway enhances sorafenib sensitivity of HepG2 cells
- Journal of Third Military Medical University Vol. 42, Issue 22, Pages: 2195-2201(2020)
- 作者机构：
  
  400038 重庆，陆军军医大学(第三军医大学)基础医学院生物化学与分子生物学教研室
- 作者简介：
- 基金信息：
  
  the General Program of National Natural Science Foundation of China(81572375);the General Program of Natural Science Foundation of Chongqing(CSTC2018jcyjA2018)
- DOI：10.16016/j.1000-5404.202007062
  CLC： R73-362;R735.7;R979.14
- Received：08 July 2020，
  
  Revised：2020-09-18，
  
  Published：30 November 2020
- 稿件说明：
移动端阅览
Yueting ZHANG, Hanxi XIAO, Liangbo SUN, et al. Activation of miR-101-3p/EZH2 pathway enhances sorafenib sensitivity of HepG2 cells[J]. Journal of Third Military Medical University, 2020, 42(22): 2195-2201.
DOI：

Yueting ZHANG, Hanxi XIAO, Liangbo SUN, et al. Activation of miR-101-3p/EZH2 pathway enhances sorafenib sensitivity of HepG2 cells[J]. Journal of Third Military Medical University, 2020, 42(22): 2195-2201. DOI： 10.16016/j.1000-5404.202007062.

摘要

目的

2

探讨激活miR-101-3p/EZH2通路在增敏索拉非尼抗肝癌HepG2细胞中的意义。

方法

2

将人肝癌细胞HepG2分成对照组(DMSO)和索拉非尼处理组(10 μmol/L)，处理24 h后，定量PCR检测miR-101-3p表达水平变化；生物信息学预测miR-101-3p结合的靶基因；在肝癌细胞中过表达miR-101-3p后Western blot和定量PCR检测zeste同源蛋白2增强子(enhancer of zeste homolog 2 protein，EZH2)表达水平变化；双荧光素酶报告基因实验验证miR-101-3p和EZH2的靶向关系；Western blot检测索拉非尼处理对EZH2表达水平的影响；在HepG2细胞中过表达miR-101-3p或添加EZH2抑制剂EPZ-6438后联合索拉非尼处理，CCK-8检测对HepG2细胞存活的影响。

结果

2

索拉非尼处理后HepG2细胞中miR-101-3p表达水平显著降低(

P

<

0.05)；miR-101-3p可在HepG2细胞中下调EZH2的表达(

P

<

0.05)；miR-101-3p与EZH2 mRNA 3’-UTR存在直接结合(

P

<

0.05)；索拉非尼处理后HepG2细胞中EZH2表达水平显著升高(

P

<

0.05)；过表达miR-101-3p或EZH2抑制剂EPZ-6438均可增加索拉非尼对HepG2细胞杀伤的敏感性(

P

<

0.05)。

结论

2

激活miR-101-3p/EZH2通路可增敏索拉非尼的抗肝癌细胞HepG2效果。

Abstract

Objective

2

To investigate the effect of activating the miR-101-3p/EZH2 pathway on sorafenib sensitivity of HepG2 cells.

Methods

2

HepG2 cells treated with sorafenib (10 μmol/L) for 24 h were examined for protein expression of enhancer of zeste homolog 2 (EZH2) using Western blotting and for mRNA expression of EZH2 and miR-101-3p using RT-qPCR. The interaction between miR-101-3p and EZH2 mRNA was analyzed through informatics analysis and verified using dual luciferase reporter assay. HepG2 cells were transfected with miR-101-3p mimic or treated with the EZH2 inhibitor EPZ-6438 followed by treatment with sorafenib for 24 h

and the change in cell proliferation was assessed using CCK-8 assay.

Results

2

Sorafenib obviously decreased the expression of miR-101-3p in HepG2 cells (

P

<

0.05). Informatics analysis showed that miR-101-3p could bind to the 3'-UTR of EZH2 mRNA

and transfection with miR-101-3p significantly inhibited the expression of EZH2 in HepG2 cells (

P

<

0.05). Sorafenib treatment significantly increased the expression of EZH2 in HepG2 cells (

P

<

0.05). Both miR-101-3p over-expression and EPZ-6438 treatment resulted in significantly increased sorafenib sensitivity of HepG2 cells (

P

<

0.05).

Conclusion

2

Activation of the miR-101-3p/EZH2 pathway can enhance sorafenib sensitivity of HepG2 cells.

关键词

Keywords

references

YANG J D, HAINAUT P, GORES G J, et al . A global view of hepatocellular carcinoma: trends, risk, prevention and management[J ] . Nat Rev Gastroenterol Hepatol, 2019, 16(10): 589-604. DOI: 10.1038/s41575-019-0186-y.

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.

LLOVET J M, VILLANUEVA A, LACHENMAYER A, et al . Advances in targeted therapies for hepatocellular car-cinoma in the genomic era[J ] . Nat Rev Clin Oncol, 2015, 12(7): 408-424. DOI: 10.1038/nrclinonc.2015.103.

COLAGRANDE S, INGHILESI A L, ABURAS S, et al . Challenges of advanced hepatocellular carcinoma[J ] . World J Gastroenterol, 2016, 22(34): 7645. DOI: 10.3748/wjg.v22.i34.7645.

WANG P, TAN G, ZHU M, et al . Hand-foot skin reaction is a beneficial indicato r of sorafenib therapy for patients with hepatocellular carcinoma: a systemic review and meta-analysis[J ] . Expert Rev Gastroenterol Hepatol, 2018, 12(1): 1-8. DOI: 10.1080/17474124.2017.1373018.

MICHLEWSKI G, CÁCERES J F. Post-transcriptional control of miRNA biogenesis[J]. RNA, 2019, 25(1): 1-16. DOI: 10.1261/rna.068692.118.

ACUNZO M, ROMANO G, WERNICKE D, et al . MicroRNA and cancer-a brief overview[J ] . Adv Biol Regul, 2015, 57(1): 1-9. DOI: 10.1016/j.jbior.2014.09.013.

ZHANG H F, WANG Y C, HAN Y D. MicroRNA-34a inhibits liver cancer cell growth by reprogramming glucose meta-bolism[J]. Mol Med Rep, 2018, 17(3): 4483-4489. DOI: 10.3892/mmr.2018.8399.

GOMES B C, RUEFF J, RODRIGUES A S. MicroRNAs and cancer drug resistance[M]Methods in Molecular Biology. New York, NY: Springer New York, 2016: 137-162. DOI: 10.1007/978-1-4939-3347-1_9.

YAN S, SHAN X, CHEN K, et al . LINC00052/miR-101-3p axis inhibits cell proliferation and metastasis by targeting SOX9 in hepatocellular carcinoma[J ] . Gene, 2018, 679(6): 138-149. DOI: 10.1016/j.gene.2018.08.038.

CAO S, LIN L, XIA X, et al . lncRNA SPRY4-IT1 regulates cell proliferation and migration by sponging miR-101-3p and regulating AMPK expression in gastric cancer[J ] . Mol Ther Nucleic Acids, 2019, 17: 455-464. DOI: 10.1016/j.omtn.2019.04.030.

XUE M Y, CAO H X. Long non-coding RNA CASC15 promotes nasopharyngeal carcinoma cell proliferation and metastasis by downregulating miR-101-3p[J]. Eur Rev Med Pharmacol Sci, 2019, 23(20): 8897-8904. DOI: 10.26355/eurrev_201910_19285.

LIU D, LI Y, LUO G, et al . LncRNA SPRY4-IT1 sponges miR-101-3p to promote proliferation and metastasis of bladder cancer cells through up-regulating EZH2[J ] . Cancer Lett, 2017, 388: 281-291. DOI: 10.1016/j.canlet.2016.12.005.

ZHANG X F, GAO D F, FANG K, et al . Med19 is targeted by miR-101-3p/miR-422a and promotes breast cancer progression by regulating the EGFR/MEK/ERK signaling pathway[J ] . Cancer Lett, 2019, 444: 105-115. DOI: 10.1016/j.canlet.2018.12.008.

VENOOK A P, PAPANDREOU C, FURUSE J, et al . The incidence and epidemiology of hepatocellular carcinoma: a global and regional perspective[J ] . Oncologist, 2010, 15(Suppl 4): 5-13. DO I: 10.1634/theoncologist.2010-s4-05.

XIE L, ZENG Y, DAI Z, et al . Chemical and genetic inhibition of STAT3 sensitizes hepatocellular carcinoma cells to sorafenib induced cell death[J ] . Int J Biol Sci, 2018, 14(5): 577-585. DOI: 10.7150/ijbs.22220.

SHENG J, QIN H, ZHANG K, et al . Targeting autophagy in chemotherapy-resistant of hepatocellular carcinoma[J ] . Am J Cancer Res, 2018, 8(3): 354-365.

KLUNGBOONKRONG V, DAS D, MCLENNAN G. Mole-cular mechanisms and targets of therapy for hepatocellular carcinoma[J]. J Vasc Interv Radiol, 2017, 28(7): 949-955. DOI: 10.1016/j.jvir.2017.03.002.

SARTORIUS K, SATORIUS B, WINKLER C, et al .The biological and diagnostic role of miRNA's in hepatocellular carcinoma[J ] . Front Biosci (Landmark Ed), 2018, 23: 1701-1720. DOI: 10.2741/4668.

QADIR M I, FAHEEM A. miRNA: a diagnostic and therapeutic tool for pancreatic cancer[J]. Crit Rev Eukaryot Gene Expr, 2017, 27(3): 197-204. DOI: 10.1615/critreveukaryotgeneexpr.2017019494.

MASOOD Y, KQUEEN C Y, RAJADURAI P. Role of miRNA in head and neck squamous cell carcinoma[J]. Expert Rev Anticancer Ther, 2015, 15(2): 183-197. DOI: 10.1586/14737140.2015.978294.

HE C, DONG X, ZHAI B, et al . MiR-21 mediates sorafenib resistance of hepatocellular carcinoma cells by inhibiting autophagy via the PTEN/Akt pathway[J ] . Oncotarget, 2015, 6(30): 28867-28881. DOI: 10.18632/oncotarget.4814.

XU Y, HUANG J, MA L, et al . MicroRNA-122 confers sorafenib resistance to hepatocellular carcinoma cells by targeting IGF-1R to regulate RAS/RAF/ERK signaling pathways[J ] . Cancer Lett, 2016, 371(2): 171-181. DOI: 10.1016/j.canlet.2015.11.034.

YANG F, LI Q J, GONG Z B, et al . MicroRNA-34a targets Bcl-2 and sensitizes human hepatocellular carcinoma cells to sorafenib treatment[J ] . Technol Cancer Res Treat, 2014, 13(1): 77-86. DOI: 10.7785/tcrt.2012.500364.

MU W, HU C, ZHANG H, et al . MiR-27b synergizes with anticancer drugs via p53 activation and CYP1B1 suppression[J ] . Cell Res, 2015, 25(4): 477-495. DOI: 10.1038/cr.2015.23.

WANG C, LIU B. MiR-101-3p induces autophagy in endometrial carcinoma cells by targeting EZH2[J]. Arch Gynecol Obstet, 2018, 297(6): 1539-1548. DOI: 10.1007/s00404-018-4768-7.

LI L, SHAO M Y, ZOU S C, et al . MiR-101-3p inhibits EMT to attenuate proliferation and metastasis in glioblastoma by targeting TRIM44[J ] . J Neuro-Oncol, 2019, 141(1): 19-30. DOI: 10.1007/s11060-018-2973-7.

ZHANG X Q, HE X J, LIU Y B, et al . MiR-101-3p inhibits the growth and metastasis of non-small cell lung cancer through blocking PI3K/AKT signal pathway by targeting MALAT-1[J ] . Biomed Pharmacother, 2017, 93: 1065-1073. DOI: 10.1016/j.biopha.2017.07.005.

SU M, XIAO Y, TANG J, et al . Role of lncRNA and EZH2 interaction/regulatory network in lung cancer[J ] . J Cancer, 2018, 9(22): 4156-4165. DOI: 10.7150/jca.27098.

LI B, CHNG W J. EZH2 abnormalities in lymphoid malignancies: underlying mechanisms and therapeutic implications[J]. J Hematol Oncol, 2019, 12(1): 118. DOI: 10.1186/s13045-019-0814-6.

NING X, SHI Z, LIU X, et al . DNMT1 and EZH2 mediated methylation silences the microRNA-200b/a/429 gene and promotes tumor progression[J ] . Cancer Lett, 2015, 359(2): 198-205. DOI: 10.1016/j.canlet.2015.01.005.

HUANG B, HUANG M, LI Q. MiR-137 suppresses migration and invasion by targeting EZH2-STAT3 signaling in human hepatocellular carcinoma[J]. Pathol Res Pract, 2018, 214(12): 1980-1986. DOI: 10.1016/j.prp.2018.08.005.

HU J, ZHANG J, SUN F, et al . Enhancer of zeste 2 polycomb repressive complex 2 subunit promotes sorafenib resistance of hepatocellular carcinoma though insulin-like growth factor 1 receptor[J ] . Anticancer Drugs, 2019, 30(7): e0746. DOI: 10.1097/cad.0000000000000746.

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