IL-17A对肝癌细胞系干性的影响

李可心; 臧梦雅; 王冬梅; 贺欢; 王宇婷; 曲春枫

doi:10.3969/j.issn.1001-5256.2017.06.023

IL-17A对肝癌细胞系干性的影响

DOI: 10.3969/j.issn.1001-5256.2017.06.023

1. 国家癌症中心/中国医学科学院北京协和医学院肿瘤医院免疫学研究室
2. 国家癌症中心/中国医学科学院北京协和医学院肿瘤医院分子肿瘤学国家重点实验室

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中图分类号: R735.7
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出版历程
- 出版日期: 2017-06-20

Effect of interleukin-17A on stemness of hepatoma cell lines

摘要

摘要: 目的探讨IL-17A对人肝癌细胞系Hep 3B、MHCC97H及MHCC97L干性的影响,以期分析IL-17A与肝癌发展之间的关系。方法选用人肝癌细胞系Hep 3B、MHCC97H及MHCC97L通过体外3D成球实验分析IL-17A对其成球能力的影响,分为只含有普通培养液的对照组和加50 ng/ml IL-17A的实验组。采用实时无标记动态细胞分析技术检测IL-17A对成球能力增强的肝癌细胞增殖及迁移的影响;采用实时定量PCR检测分析IL-17A对成球能力增强的肝癌细胞所表达的IL-17A受体IL-17RA、IL-17RC以及干性相关基因SOX2、NANOG、OCT4、BMI1的mRNA表达的影响;通过Western Blot检测上皮-间质转换（EMT）相关蛋白E-钙黏蛋白（E-cadherin）、N-钙黏蛋白（N-cadherin）、波形蛋白（vimentin）的表达。计量资料组间比较采用t检验。结果在存在50 ng/ml IL-17A及接种细胞数均为500个的条件下,Hep 3B肝癌细胞的成球数目显著增加[由（113.0±10.3）个增加至（180.0±7.2）个,t=5.533,...
- 肝肿瘤,实验性 /
- 细胞系,肿瘤 /
- 白细胞介素17 /
- 肿瘤干细胞
Abstract: Objective To investigate the effect of interleukin-17A (IL-17A) on stemness of human hepatoma cell lines Hep 3B, MHCC97 H, and MHCC97 L and the association between IL-17 A and the progression of liver cancer. Methods Human hepatoma cell lines Hep 3B, MHCC97 H, and MHCC97 L were selected, and in vitro 3D sphere formation assay was used to analyze the effect of IL-17 A on sphere formation ability. The control group with common culture solution and the experimental group with 50 ng/ml IL-17 A were established. Real-time cellular analysis was used to determine the effect of IL-17 A on the proliferation and migration of hepatoma cells with enhanced sphere formation ability; quantitative real-time PCR was used to measure the changes in the mRNA expression of IL-17 A receptors IL-17 RA and IL-17 RC and stemness-related genes SOX2, NANOG, OCT4, and BMI1 in hepatoma cells with enhanced sphere formation ability; Western blot was used to measure the expression of epithelial-mesenchymal transition-related proteins E-cadherin, N-cadherin, and vimentin. The t-test was used for comparison of continuous doota betwwen groups. Results With the presence of 50 ng/ml IL-17 A and 500 inoculated cells, Hep 3B cells had a significant increase in the number of spheres formed (113. 0 ± 10. 3 vs 180. 0 ± 7. 2, t =5. 533, P < 0. 001) , while MHCC97 H and MHCC97 L cells showed no significant changes (t = 1. 087 and 0. 279, P = 0. 325 and 0. 785) .The analysis showed that IL-17 A promoted the proliferation and migration of Hep 3B cells with an increased number of spheres formed. After the addition of 50 ng/ml IL-17 A, there was an increase in the mRNA expression of IL-17 A receptors IL-17 RA and IL-17 RC over the time of treatment; Hep 3B cells showed significant increases in the mRNA expression of stemness-related genes SOX2 (t = 4. 749, P =0. 042) , NANOG (t = 19. 600, P = 0. 003) , OCT4 (t = 37. 310, P < 0. 001) , and BMI1 (t = 16. 810, P = 0. 004) . Western blot showed no significant change in the expression of the epithelium-derived marker E-Cadherin; there was an increase in the expression of the interstitial marker N-cadherin after the treatment with 50 ng/ml IL-17 A for 48 hours (t = 17. 620, P = 0. 036) ; there was no significant change in the expression of vimentin. Conclusion To a certain degree, IL-17 A enhances the stemness of Hep 3B hepatoma cells.
- liver neoplasms, experimental /
- cell line, tumor /
- interleukin-17 /
- neoplastic stem cells

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参考文献(17)

[1]HERNANDEZ-GEA V, TOFFANIN S, FRIEDMAN SL, et al.Role of the microenvironment in the pathogenesis and treatment of hepatocellular carcinoma[J].Gastroenterology, 2013, 144 (3) :512-527.

[2]BUDHU A, FORGUES M, YE QH, et al.Prediction of venous metastases, recurrence, and prognosis in hepatocellular carcinoma based on a unique immune response signature of the liver microenvironment[J].Cancer cell, 2006, 10 (2) :99-111.

[3]MOSELEY TA, HAUDENSCHILD DR, ROSE L, et al.Interleukin-17 family and IL-17 receptors[J].Cytokine Growth Factor Rev, 2003, 14 (2) :155-174.

[4]TOY D, KUGLER D, WOLFSON M, et al.Cutting edge:interleukin 17 signals through a heteromeric receptor complex[J].J Immunol, 2006, 177 (1) :36-39.

[5]WANG M, WANG L, REN T, et al.IL-17A/IL-17RA interaction promoted metastasis of osteosarcoma cells[J].Cancer Biol Ther, 2013, 14 (2) :155-163.

[6]WANG K, KIM MK, DI CARO G, et al.Interleukin-17 receptor a signaling in transformed enterocytes promotes early colorectal tumorigenesis[J].Immunity, 2014, 41 (6) :1052-1063.

[7]WU J, DU J, LIU L, et al.Elevated pretherapy serum IL-17 in primary hepatocellular carcinoma patients correlate to increased risk of early recurrence after curative hepatectomy[J].PLo S One, 2012, 7 (12) :e50035.

[8]BANDHAVKAR S.Cancer stem cells:a metastasizing menace![J].Cancer Med, 2016, 5 (4) :649-655.

[9]BAO S, WU Q, MCLENDON RE, et al.Glioma stem cells promote radioresistance by preferential activation of the DNA damage response[J].Nature, 2006, 444 (7120) :756-760.

[10]CHIBA T, KAMIYA A, YOKOSUKA O, et al.Cancer stem cells in hepatocellular carcinoma:Recent progress and perspective[J].Cancer Lett, 2009, 286 (2) :145-153.

[11]HONG S, SONG HR, LUTZ K, et al.Type II protein arginine methyltransferase 5 (PRMT5) is required for circadian period determination in Arabidopsis thaliana[J].Proc Natl Acad Sci U S A, 2010, 107 (49) :21211-21216.

[12]ISHIKAWA M, YOSHITOMI T, ZORUMSKI CF, et al.Downregulation of glutamine synthetase via GLAST suppression induces retinal axonal swelling in a rat ex vivo hydrostatic pressure model[J].Invest Ophthalmol Vis Sci, 2011, 52 (9) :6604-6616.

[13]LIU J, TAN Y, ZHANG H, et al.Soft fibrin gels promote selection and growth of tumorigenic cells[J].Nat Mater, 2012, 11 (8) :734-741.

[14]KNOWLES BB, HOWE CC, ADEN DP.Human hepatocellular carcinoma cell lines secrete the major plasma proteins and hepatitis B surface antigen[J].Science, 1980, 209 (4455) :497-499.

[15]TIAN J, TANG ZY, YE SL, et al.New human hepatocellular carcinoma (HCC) cell line with highly metastatic potential (MHCC97) and its expressions of the factors associated with metastasis[J].Br J Cancer, 1999, 81 (5) :814-821.

[16]LI Y, TANG ZY, YE SL, et al.Establishment of cell clones with different metastatic potential from the metastatic hepatocellular carcinoma cell line MHCC97[J].World J Gastroenterol, 2001, 7 (5) :630-636.

[17]KANG Y, MASSAGUE J.Epithelial-mesenchymal transitions:twist in development and metastasis[J].Cell, 2004, 118 (3) :277-279.

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