SOX11 抑制食管癌细胞增殖和迁移的机制研究

doi:10.11904/j.issn.1002-3070.2020.01.004

摘要/Abstract

摘要： 目的探讨转录因子SOX11 在食管癌组织和细胞系中的表达及对人食管癌细胞EC109增殖和迁移的影响,并初步探讨其作用机制。方法 qRT-PCR和免疫组织化学法分析SOX11在食管癌组织、细胞系(EC109、KYSE150、KYSE410 和 KYSE510)及正常食管上皮细胞 HEEC的表达;分别转染 pcDNA3.1(+)-Flag-SOX11 质粒(实验组)和pcDNA3.1(+)-Flag-Vector质粒(对照组)至EC109细胞系,qRT-PCR验证SOX11的过表达;CCK-8实验和Transwell实验分析SOX11对细胞增殖和迁移能力的影响;qRT-PCR及Western blot验证SOX11对细胞增殖和迁移的相关标志物表达分析。结果与食管上皮组织相比,SOX11 mRNA 和蛋白在食管癌组织中的表达明显下调;SOX11的蛋白表达下调与肿瘤分级(P=0.014)、T分期(P=0.036)、淋巴结转移(P=0.014)相关;同时与食管上皮细胞相比,SOX11 mRNA在食管癌细胞中表达下调,差异具有统计学意义(P＜0.001);与对照组相比,实验组24、48、72 h细胞活性受到明显抑制(P＜0.05);实验组细胞迁移能力明显受到抑制(P＜0.001);与对照组相比,实验组中active-β-catenin 及下游的基因受到了明显的抑制。结论 SOX11在人食管癌组织和细胞系中表达下调,可能通过拮抗Wnt/β-catenin信号通路参与食管癌的进程。

关键词: 食管癌, SOX11, 抑癌基因, 增殖和迁移, Wnt/β-catenin

Abstract: Objective The aim of this study was to investigate the expression of the transcription factor SOX11 in esophageal cancer tissues and cell lines,the effects on proliferation and migration of esophageal cancer EC109 cells,and to explore its mechanism of action.Methods RT-qPCR and immunohistochemistry were used to analyze the expression of SOX11 in esophageal tissues and cell lines;SOX11 plasmids were transfected into low expression esophageal cancer cells(EC109,KYSE150,KYSE410 and KYSE510)and normal esophageal epithelial HEEC cells.The overexpression of SOX11 was confirmed in EC109 cells transfected with pcDNA3.1(+)-Flag-SOX11(the experimental group)and pcDNA3.1(+)-Flag-vector plasmid(the control group).CCK-8 and Transwell assays were used to analyze the effects of SOX11 on proliferation and migration of EC109 cells.RT-qPCR and Western blot assays were used to verify the effect of SOX11 on cell proliferation and migration of EC109 cells.Results Compared with esophageal epithelial tissues,the expression of SOX11 at levels of mRNA and protein was significantly down-regulated in esophageal cancer tissues;the down-regulated expression of SOX11 protein was associated with tumor grade(P=0.014),T stage(P=0.036)and lymph node metastasis(P=0.014).At the same time,compared with esophageal epithelial cells,the mRNA expression of SOX11 was down-regulated in esophageal cancer cells;the difference was statistically significant(P＜0.001).Compared with the control group(empty vector),the cell viability was significantly suppressed in EC109 cells after transfected with SOX11 mimics for 24,48 and 72 h(P＜0.05);the cell migration ability of EC109 cells was significantly inhibited in the experimental group(P＜0.001);Compared with the control group,active-β-catenin and its downstream genes were also significantly suppressed in the experimental group.Conclusion SOX11 is down-regulated in human esophageal cancer tissues and cancer cell lines,and may participate in the process of esophageal cancer by antagonizing the Wnt/β-catenin signaling pathway.

Key words: Esophageal cancer, SOX11, Tumor suppressor genes, Cell proliferation/migration, Wnt/β-catenin

中图分类号:

R735

赵楠楠, 秦文, 韩永焕, 鲁静, 李亚敏, 徐亚君. SOX11 抑制食管癌细胞增殖和迁移的机制研究[J]. 实用肿瘤学杂志, 2020, 34(1): 17-23.

ZHAO Nannan, QIN Wen, HAN Yonghuan, LU Jing, LI Yamin, XU Yajun. Effects of SOX11 on inhibiting proliferation and migration of esophageal cancer cells[J]. Journal of Practical Oncology, 2020, 34(1): 17-23.

参考文献

1 Qiu ZH,Zhang WW,Zhang HH,et al.Brucea javanica oil emulsion improves the effect of radiotherapy on esophageal cancer cells by inhibiting cyclin D1-CDK4/6 axis[J].World J Gastroenterol,2019,25(20):2463-2472.
2 Huang J,Liu H,Sun T,et al.Omeprazole prevents CDX2 and SOX9 expression by inhibiting hedgehog signaling in Barrett′s esophagus cells[J].Clin Sci(Lond),2019,133(3):483-495.
3 张茂祥,孙惠昕,宋冰冰.2008-2012年哈尔滨市道里区恶性肿瘤发病与死亡分析[J].实用肿瘤学杂志,2019,33(3):261-265.
4 Xu S,Yu J,Wang Z,et al.SOX11 promotes osteoarthritis through induction of TNF-α[J].Pathol Res Pract,2019,6(2):152442-152451.
5 Qin Y,Sun B,Zhang F,et al.Sox7 is involved in antibody-dependent endothelial cell activation and renal allograft injury via the Jagged1-Notch1 pathway[J].Exp Cell Res,2019,375(2):20-27.
6 Wurm A,Sock E,Fuchshofer R,et al.Anterior segment dysgenesis in the eyes of mice deficient for the high-mobility-group transcription factor Sox11[J].Exp Eye Res,2008,86(6):895-907.
7 Dictor M,Ek S,Sundberg M,et al.Strong lymphoid nuclear expression of SOX11 transcription factor defines lymphoblastic neoplasms,mantle cell lymphoma and Burkitt′s lymphoma[J].Haematologica,2009,94(11):1563-1568.
8 Hide T,Takezaki T,Nakatani Y,et al.Sox11 prevents tumorigenesis of glioma-initiating cells by inducing neuronal differentiation[J].Cancer Res,2009,69(20):7953-7959.
9 Petrakis G,Veloza L,Clot G,et al.Increased tumor angiogenesis in SOX11-positive mantle cell lymphoma[J].Histopathology,2019,7(2):281-291.
10 Xu J,Wang L,Li J,et al.SOX11-negative mantle cell lymphoma:clinicopathologic and prognostic features of 75 patients[J].Am J Surg Pathol,2019,43(5):710-716.
11 Jian J,Wang GY,Zhao J.Increased expression of sex determining region Y-box 11(SOX11)in cutaneous malignant melanoma[J].J Int Med Res,2013,41(4):1221-1227.
12 Shepherd JH,Uray IP,Mazumdar A,et al.The SOX11 transcription factor is a critical regulator of basal-like breast cancer growth,invasion,and basal-like gene expression[J].Oncotarget,2016,7(11):13106-13121.
13 Xu S,Dong Y,Huo Z,et al.SOX11:a potentially useful marker in surgical pathology:a systematic analysis of SOX11 expression in epithelial and non-epithelial tumours[J].Histopathology,2019,74(3):391-405.
14 Tosic N,Petrovic I,Grujicic NK,et al.Prognostic significance of SOX2,SOX3,SOX11,SOX14 and SOX18 gene expression in adult de novo acute myeloid leukemia[J].Leuk Res,2018,67(3):32-38.
15 Palomero J,Vegliante MC,Eguileor A,et al.SOX11 defines two different subtypes of mantle cell lymphoma through transcriptional regulation of BCL6[J].Leukemia,2016,30(7):1596-1599.
16 National Health Commission of The People′s Republic of China.Chinese guidelines for diagnosis and treatment of esophageal carcinoma 2018(English version)[J].Chin J Cancer Res,2019,31(2):223-258.
17 Fatima I,El-Ayachi I,Taotao L,et al.The natural compound Jatrophone interferes with Wnt/β-catenin signaling and inhibits proliferation and EMT in human triple-negative breast cancer[J].PLoS One,2017,12(12):e0189864.
18 Li C,Lyu J,Meng QH.MiR-93 Promotes Tumorigenesis and metastasis of non-small cell lung cancer cells by activating the PI3K/Akt pathway via inhibition of LKB1/PTEN/CDKN1A[J].J Cancer,2017,8(5):870-879.
19 You A,Fokas E,Wang LF,et al.Expression of the Wnt antagonist DKK3 is frequently suppressed in sporadic epithelial ovarian cancer[J].J Cancer Res Clin Oncol,2011,137(4):621-627.
20 Zhang S,Li S,Gao JL.Promoter methylation status of the tumor suppressor gene SOX11 is associated with cell growth and invasion in nasopharyngeal carcinoma[J].Cancer Cell Int,2013,13(1):109.
21 Soldini D,Valera A,Solé C,et al.Assessment of SOX11 expression in routine lymphoma tissue sections:characterization of new monoclonal antibodies for diagnosis of mantle cell lymphoma[J].Am J Surg Patho,2014,38(1):86-93.
22 Lu TX,Li JY,Xu W.The role of SOX11 in mantle cell lymphoma[J].Leuk Res,2013,37(11):1412-1419.
23 Gadi J,Jung SH,Lee MJ,et al.The transcription factor protein Sox11 enhances early osteoblast differentiation by facilitating proliferation and the survival of mesenchymal and osteoblast progenitors[J].J Biol Chem,2013,288(35):25400-25413.
24 Cizelsky W,Hempel A,Metzig M,et al.sox4 and sox11 function during Xenopus laevis eye development[J].PLoS One,2013,8(7):e69372.
25 Meggendorfer M,Kern W,Haferlach C,et al.SOX11 overexpression is a specific marker for mantle cell lymphoma and correlates with t(11;14)translocation,CCND1 expression and an adverse prognosis[J].Leukemia,2013,27(12):2388-23891.

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[4]	于芳, 赵培. TRAP1在食管癌发展中的作用及机制研究[J]. 实用肿瘤学杂志, 2018, 32(6): 515-519.
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[6]	靳晶综述，贺宇彤审校. miR-21在食管癌中的研究进展[J]. 实用肿瘤学杂志, 2017, 31(3): 277-280.
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[11]	薛伟男,张干,杨艳梅,马玉彦,孙根,崔滨滨. 抑癌基因ING1在大肠癌中的表达及预后意义[J]. 实用肿瘤学杂志, 2014, 28(5): 385-390.
[12]	顾红媛综述樊立华审校. 宫颈癌致病危险因素的研究进展[J]. 实用肿瘤学杂志, 2014, 28(4): 367-370.
[13]	王海峰,曲颜丽,田含含,王义海,曾敏. CYP1A1、GSTM1基因多态性及环境因素与新疆哈萨克族食管癌家族聚集性的相关性研究[J]. 实用肿瘤学杂志, 2013, 27(5): 385-390.
[14]	赵尧平综述,姜丽华审校. 胸段硬膜外麻醉对食管癌术后心律失常的影响[J]. 实用肿瘤学杂志, 2013, 27(3): 282-284.
[15]	马臣,姜永晓,全培良,刘曙正,陆建邦,陈,琼,孙喜斌. 河南省居民1985—2009年食管癌死亡率分析及预测[J]. 实用肿瘤学杂志, 2013, 27(2): 124-129.