LncRNA-NEF转录激活FOXA2并通过抑制β-catenin信号通路抑制肝癌细胞的EMT过程

肝脏 ›› 2021, Vol. 26 ›› Issue (10): 1137-1141.

LncRNA-NEF转录激活FOXA2并通过抑制β-catenin信号通路抑制肝癌细胞的EMT过程

车军, 杨柳青, 孙斌, 贾泽博

712000 陕西咸阳市中心医院消化内科(车军,贾泽博);西安医学院第二附属医院消化科(杨柳青);汉中市中心医院消化内科内镜中心(孙斌)

收稿日期:2020-11-27 出版日期:2021-10-31 发布日期:2021-12-07
通讯作者: 孙斌,Email:362610078@qq.com
基金资助:
咸阳市科技局项目(16K69-02)

LncRNA-NEF transcriptionally activates FOXA2 and inhibits epithelial–mesenchymal transition of hepatocellular carcinoma cells by inhibiting β-catenin signaling pathway

CHE Jun¹, YANG Liu-qing², SUN Bin³, JIA Ze-bo¹

1. Department of Gastroenterology, Xianyang Central Hospital, Shaanxi 712000, China;
2. Department of Gastroenterology, The Second Affiliated Hospital of Xi'an Medical College, Shaanxi 710038, China;
3. Endoscopy Center of Gastroenterology, Hanzhong Central Hospital, Shaanxi 723000, China

Received:2020-11-27 Online:2021-10-31 Published:2021-12-07
Contact: Sun Bin, Email: 362610078@qq.com

摘要/Abstract

摘要： 目的探讨长链非编码lncRNA-NEF对肝癌转移关键步骤EMT过程的分子调节机制。方法通过TGF-β诱导肝癌细胞系Hep3B细胞向EMT转化,通过构建pcDNA3.1-FOXA2过表达载体,过表达FOXA2诱导肝癌细胞系HepG2细胞向MET转化,随后检测两种细胞的E-cadherin、Vimentin、Snail共3种EMT相关标志物的表达变化、lncRNA-NEF的表达变化,以及细胞侵袭能力变化。之后检测上述两种细胞分别经历EMT和MET转化后,β-catenin信号通路相关蛋白因子的表达变化情况。结果 lncRNA-NEF在肝癌组织和肝癌细胞中表达水平低于对照组正常细胞(P<0.05);与对照组相比,TGF-β处理组的细胞侵袭能力显著增强(P<0.05),pcDNA3.1-FOXA2转染组的细胞侵袭能力显著降低(P<0.05);与对照组相比,TGF-β处理组的Hep3B细胞内磷酸化的β-catenin(p-β-catenin)表达量显著增高,而总β-catenin蛋白水平不变,pcDNA3.1-FOXA2转染组的HepG2细胞内磷酸化的β-catenin(p-β-catenin)表达量显著降低,而总β-catenin蛋白水平不变。LncRNA-NEF是以顺式作用方式转录激活FOXA2,并通过抑制β-catenin信号通路抑制肝癌细胞的EMT过程。结论 LncRNA-NEF能转录激活FOXA2并通过抑制β-catenin信号通路抑制肝癌细胞的EMT过程。

关键词: lncRNA-NEF, FOXA2, EMT, MET, β-catenin信号通路

Abstract: Objective To explore the molecular mechanism of long-chain non-coding RNA-neighboring enhancer of FOXA2 (lncRNA-NEF) on the epithelial-mesenchymal transition (EMT) process of hepatocellular carcinoma (HCC), a key step in liver cancer metastasis. Methods HCC line Hep3B was treated with transforming growth factor-β (TGF-β) to induce EMT. A pcDNA3.1-FOXA2 overexpression vector was constructed and transduced into another HCC line HepG2 to overexpress FOXA2 and induce mesenchymal-epithelial transition (MET). The EMT-related markers including E-cadherin, Vimentin and Snail were detected and cell invasion assay was performed in these lines with and without relative treatment. The expression of lncRNA-NEF was determined in 10 paired operated liver cancer tissues and adjacent normal tissues, as well as in the HCC lines. Subsequently, the protein expression of factors related to β-catenin signaling pathway before and after EMT or MET transformation in the HCC lines were detected. Results The expression level of lncRNA-NEF in liver cancer tissues and HCC lines was lower than that in normal controls (P<0.05). Compared with the control cells, the cell invasion ability of TGF-β treated HCC cells significantly enhanced (P<0.05), but the cell invasion ability of PCDNA3.1-FOXA2 transduced cells significantly reduced (P<0.05). The expression level of phosphorylated β-catenin (p-β-catenin) in Hep3B cells treated with TGF-β increased significantly when compared with the control cells. On the contrary, the expression level of phosphorylated β-catenin (P-β-catenin) in HepG2 cells transfected with PCDNA3.1-FoxA2 significantly decreased. The level of total β-catenin protein remained unchanged in both HCC lines with and without relative treatment. LncRNA-NEF activated FOXA2 in a cis-acting manner, and inhibited the EMT process of liver cancer cells by inhibiting the β-catenin signaling pathway. Conclusion LncRNA-NEF activates FOXA2 transcription and inhibits the EMT process of liver cancer cells by inhibiting the β-catenin signaling pathway.

Key words: LncRNA-NEF, FOXA2, EMT, MET, β-catenin signaling pathway

车军, 杨柳青, 孙斌, 贾泽博. LncRNA-NEF转录激活FOXA2并通过抑制β-catenin信号通路抑制肝癌细胞的EMT过程[J]. 肝脏, 2021, 26(10): 1137-1141.

CHE Jun, YANG Liu-qing, SUN Bin, JIA Ze-bo. LncRNA-NEF transcriptionally activates FOXA2 and inhibits epithelial–mesenchymal transition of hepatocellular carcinoma cells by inhibiting β-catenin signaling pathway[J]. Chinese Hepatolgy, 2021, 26(10): 1137-1141.

参考文献

[1] Fitzmaurice C, Allen C, Barber RM, et al. Global, regional, and national cancer incidence, mortality, years of life lost, years lived with disability, and disability-adjusted life-years for 32 cancer groups, 1990 to 2015: A systematic analysis for the global burden of disease study. JAMA Oncol, 2017, 3:524-548.
[2] Massagué J, Batlle E, Gomis RR. Understanding the molecular mechanisms driving metastasis. Mol Oncol, 2017, 11(1):3-4.
[3] Brabletz T, Kalluri R, Nieto MA, et al. EMT in cancer.. Nat Rev Cancer, 2018, 18:128-134.
[4] Du B, Shim JS. Targeting epithelial-mesenchymal transition (EMT) to overcome drug resistance in cancer. Molecules, 2016, 21:965.
[5] Lamouille S, Xu J, Derynck R. Molecular mechanisms of epithelial–mesenchymal transition. Nat Rev Mol Cell Biol, 2014, 15:178-196.
[6] Yoshida T, Ozawa Y, Kimura T, et al. Eribulin mesilate suppresses experimental metastasis of breast cancer cells by reversing phenotype from epithelial-mesenchymal transition (EMT) to mesenchymal-epithelial transition (MET) states. Br J Cancer, 2014, 110:1497-1505.
[7] Davis FM, Stewart TA, Thompson EW, et al. Targeting EMT in cancer: opportunities for pharmacological intervention. Trends Pharmacol Sci, 2014, 35:479-488.
[8] Deng J, Wang L, Chen H, et al. Targeting epithelial-mesenchymal transition and cancer stem cells for chemoresistant ovarian cancer. Oncotarget, 2016, 7:55771-55788.
[9] Marcucci F, Stassi G, De Maria R. Epithelial-mesenchymal transition: a new target in anticancer drug discovery. Nat Rev Drug Discov, 2016, 15:311-325.
[10] Liang WC, Ren JL, Wong CW, et al. LncRNA-NEF antagonized epithelial to mesenchymal transition and cancer metastasis via cis-regulating FOXA2 and inactivating Wnt/β-catenin signaling. Oncogene, 2018, 37:1445-1456.
[11] Chen Q, Yang W, Wang X, et al. TGF-β1 Induces EMT in Bovine Mammary Epithelial Cells Through the TGFβ1/Smad Signaling Pathway. Cell Physiol Biochem, 2017, 43:82-93.
[12] Basu S, Cheriyamundath S, Ben-Ze'ev A. Cell-cell adhesion: linking Wnt/β-catenin signaling with partial EMT and stemness traits in tumorigenesis. F1000Res,2018,7:F1000 Faculty Rev-1488.

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