STAT3加重TGF-β1诱导的肝癌细胞上皮-间质转化的发生

肝脏 ›› 2018, Vol. 23 ›› Issue (2): 128-132.

STAT3加重TGF-β1诱导的肝癌细胞上皮-间质转化的发生

刘婷, 吴俊成, 陆伦根, 徐铭益

200080 上海交通大学附属第一人民医院消化科

收稿日期:2017-10-30 出版日期:2018-02-28 发布日期:2020-05-18
通讯作者: 徐铭益,Email:xumingyi2014@163.com
基金资助:
国家自然基金面上项目(81570547);国家科技部十二五项目(2012ZX10002007-001-040,2013ZX10002004-002-003);院优秀青年培养计划(061405)

STAT3 aggravates TGF-β1 induced epithelial-to-mesenchymal transition in hepatocellular carcinoma cells

LIU Ting, WU Jun-cheng, LU Lun-gen, XU Ming-yi.

Department of Gastroenterology, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200080, China

Received:2017-10-30 Online:2018-02-28 Published:2020-05-18
Contact: XU Ming-yi, Email: xumingyi2014@163.com

摘要/Abstract

摘要： 目的探讨IL-6/JAK/STAT3和Smad3/TGF-β1信号通路在肝癌细胞上皮-间质转化(EMT)发生过程中的作用。方法分别予TGF-β1、IL-6、AG490刺激人肝癌细胞株(HepG2、Bel7402、MHCC97H、HCCLM3)后,qPCR检测Snail、STAT3 的mRNA表达水平,蛋白质免疫检测p-STAT3/STAT3、p-Smad3/Smad3、TGF-β1以及EMT相关分子标志物Snail、E-Cadherin、Vimentin的蛋白表达水平。结果 TGF-β1可以诱导HepG2细胞EMT的发生,下调E-Cadherin的表达,上调Vimentin、Snai的表达。IL-6在肝癌细胞中通过刺激STAT3,激活Smad3/TGF-β1通路,进而上调EMT相关分子标志物Snail、Vimentin的表达。AG490(一种JAK2的特异性抑制剂)抑制p-STAT3/STAT3、p-Smad3及EMT相关分子标志物Snail的表达。结论 STAT3在TGF-β1诱导的肝癌细胞EMT发生过程中作为一个正向调控因子,IL6/JAK/STAT3和Snail/Smad3/TGF-β1信号通路协同促进肝癌细胞EMT的发生。

关键词: 肝细胞癌;上皮-间质转化;信号转导和转录活化蛋白3;转化生长因子-β1

Abstract: Objective To investigate the underlying molecular mechanisms of interleukin-6/Janus kinase/ signal transducer and activator of transcription (IL-6/JAK/STAT3) and Smad3/transforming growth factor-β1 (Smad3/TGF-β1) signaling pathways during the epithelial to mesenchymal transition (EMT) process in hepatocellular carcinoma (HCC) cells.Methods Human HCC cell lines HepG2 and HCCLM3 were treated with TGF-β1, AG490 and IL-6, respectively. mRNA expressions of STAT3 and Snail were measured using quantitative PCR. Expressions of p-STAT3/STAT3, p-Smad3/Smad3, TGF-β1 and EMT-related markers were measured using western blotting. Results TGF-β1 induced EMT in HepG2 cells, dampen E-Cadherin expression and up-regulate expressions of vimentin, Snail, p-Smad2/3 and p-STAT3/STAT3. IL-6 activated Smad3/TGF-β1 pathway by stimulating STAT3 in human HCC lines, and up-regulated expressions of EMT-related molecular markers (Snail and Vimentin). AG490, the JAK2-specific inhibitor, inhibited expressions of p-STAT3/STAT3, p-Smad3 and Snail.Conclusion STAT3 aggravated TGF-β1-induced EMT and HCC metastasis, which performed a synergistical effect with IL-6/JAK/STAT3 and Snail/Smad3/TGF-β1 signaling pathways.

Key words: Hepatocellular carcinoma, Epithelial-to-mesenchymal transition, Signal transducer and activator of transcription 3, Transforming growth factor-β1

刘婷, 吴俊成, 陆伦根, 徐铭益. STAT3加重TGF-β1诱导的肝癌细胞上皮-间质转化的发生[J]. 肝脏, 2018, 23(2): 128-132.

LIU Ting, WU Jun-cheng, LU Lun-gen, XU Ming-yi.. STAT3 aggravates TGF-β1 induced epithelial-to-mesenchymal transition in hepatocellular carcinoma cells[J]. Chinese Hepatolgy, 2018, 23(2): 128-132.

参考文献

[1] Zhong Z, Wen Z, Darnell JE, et al. Stat3: a STAT family member activated by tyrosine phosphorylation in response to epidermal growth factor and interleukin-6. Science, 1994, 264: 95-98.
[2] Stark GR, Jr. DJE. The JAK-STAT pathway at twenty. Immunity, 2012,36: 503-514.
[3] Yang X, Liang L, Zhang XF, et al. MicroRNA-26a suppresses tumor growth and metastasis of human hepatocellular carcinoma by targeting interleukin-6-Stat3 pathway. Hepatology, 2013,58: 158-170.
[4] Lin L, Amin R, Gallicano GI, et al. The STAT3 inhibitor NSC74859 is effective in hepatocellular cancers with disrupted TGF-b signaling. Oncogene, 2009,28:961-972.
[5] Wu K, Ding J, Chen C, et al. Hepatic transforming growth factor beta gives rise to tumor-initiating cells and promotes liver cancer development. Hepatology, 2012,56: 2255-2267.
[6] 于景霞,刘婷,李沁恺等.特异性抑制JAK/STAT3信号通路对大鼠肝细胞癌的影响.胃肠病学,2017,22: 272-275.
[7] Ding W, You H, Dang H, et al. Epithelial-to-mesenchymal transition of murine liver tumor cells promotes invasion. Hepatology,2010,52: 945-953.
[8] Nieto MA, Huang RY, Jackson RA, et al. EMT: 2016. Cell, 2016,166: 21-45.
[9] Zeisberg M, Neilson EG. Biomarkers for epithelial-mesenchymal transitions. J Clin Invest, 2009,119: 1429-1437.
[10] Liu RY, Zeng Y, Lei Z, et al. JAK/STAT3 signaling is required for TGF-beta-induced epithelial-mesenchymal transition in lung cancer cells. Int J Oncol, 2014,44: 1643-1651.
[11] Gonzalez DM, Medici D. Signaling mechanisms of the epithelial-mesenchymal transition. Sci Signal, 2014,7: re8.
[12] Batlle E, Sancho E, Francí C, et al. The transcription factor Snail is a repressor of E-cadherin gene expression in epithelial tumour cells. Nat Cell Biol, 2000,2:84-89.
[13] Horiguchi K, Shirakihara T, Nakano A, et al. Role of Ras signaling in the induction of snail by transforming growth factor-beta. J Biol Chem, 2009,284: 245-253.
[14] Sullivan NJ, Sasser AK, Axel AE, et al. Interleukin-6 induces an epithelial-mesenchymal transition phenotype in human breast cancer cells. Oncogene,2009,28: 2940-2947.