IL-17RA基因过表达对人肺腺癌A549细胞增殖、迁移和侵袭的影响
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摘要
目的:探讨白介素-17受体A(interleukin-17 receptor A,IL-17RA)对人肺腺癌A549细胞增殖、迁移及侵袭能力的影响及其可能的机制。方法:构建IL-17RA基因过表达慢病毒载体并感染A549细胞,采用RT-qPCR法和Western blot方法检测IL-17RA、MMP-2、MMP-9 mRNA和蛋白的表达。MTT实验、划痕实验及Transwell侵袭实验检测IL-17RA过表达对A549细胞增殖、迁移及侵袭的影响。结果:IL-17RA过表达A549细胞中IL-17RA mRNA和蛋白表达显著上调。IL-17RA过表达细胞的增殖活性同对照组无明显变化,但过表达组细胞的迁移距离较空载病毒组及空白对照组明显增多(P<0.05),过表达组穿膜细胞数较空载病毒组及空白对照组明显增多(P<0.05)。IL-17RA高表达可明显上调MMP-2和MMP-9蛋白的表达。结论:成功构建IL-17RA过表达A549细胞,IL-17RA具有增强A549细胞迁移及侵袭的作用,其机制可能与上调MMP-2和MMP-9表达有关。
Objective:To investigate the effect of interleukin-17 receptor A(IL-17 RA) on the proliferation,migration and invasion of lung adenocarcinoma A549 cells.Methods:A549 cells were infected with recombinant lentivirus LV-IL-17 RA.The expression of IL-17 RA,MMP-2,MMP-9 mRNA and protein were detected by Real-time qPCR and Western blot.The effect of IL-17 RA overexpression on the proliferation,migration and invasion of A549 cells were detected by MTT assay,wound healing and Transwell invasion assays.Results:The expressions of IL-17 RA mRNA and protein were stably up-regulated in the IL-17 RA overexpressed A549 cells.The proliferative activity of IL-17 RA overexpressing cells did not change significantly with the control group.However,the migration distance overexpressing cells was significantly higher than that of empty plasmid group and blank control group(P<0.05).The number of transmembrane cells in the overexpression group was significantly higher than that in the empty plasmid group and blank control group(P<0.05).High expression of IL-17 RA significantly up-regulated the expression of MMP-2 and MMP-9 proteins.Conclusion:IL-17 RA over-expressing A549 cells was successfully constructed.The overexpression of IL-17 RA may enhance the migration,and invasion of A549 cells,which may be correlated with the upregulation of MMP-2 and MMP-9.
Objective:To investigate the effect of interleukin-17 receptor A(IL-17 RA) on the proliferation,migration and invasion of lung adenocarcinoma A549 cells.Methods:A549 cells were infected with recombinant lentivirus LV-IL-17 RA.The expression of IL-17 RA,MMP-2,MMP-9 mRNA and protein were detected by Real-time qPCR and Western blot.The effect of IL-17 RA overexpression on the proliferation,migration and invasion of A549 cells were detected by MTT assay,wound healing and Transwell invasion assays.Results:The expressions of IL-17 RA mRNA and protein were stably up-regulated in the IL-17 RA overexpressed A549 cells.The proliferative activity of IL-17 RA overexpressing cells did not change significantly with the control group.However,the migration distance overexpressing cells was significantly higher than that of empty plasmid group and blank control group(P<0.05).The number of transmembrane cells in the overexpression group was significantly higher than that in the empty plasmid group and blank control group(P<0.05).High expression of IL-17 RA significantly up-regulated the expression of MMP-2 and MMP-9 proteins.Conclusion:IL-17 RA over-expressing A549 cells was successfully constructed.The overexpression of IL-17 RA may enhance the migration,and invasion of A549 cells,which may be correlated with the upregulation of MMP-2 and MMP-9.
引文
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[2] Ettinger DS,Wood DE,Aisner DL,et al.Non-small cell lung cancer,version 5.2017,NCCN clinical practice guidelines in oncology[J].J Natl Compr Canc Netw,2017,15(4):504-535.
[3] Lu M,Tian Y,Yue WM,et al.GOLPH3,a good prognostic indicator in early-stage NSCLC related to tumor angiogenesis[J].Asian Pac J Cancer Prev,2014,15(14):5793-5798.
[4] 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.
[5] Yao Z,Fanslow WC,Seldin MF,et al.Herpesvirus saimiri encodes a new cytokine,IL-17,which binds to a novel cytokine receptor[J].J Immunol,2011,187(9):4392-4402.
[6] Huang Q,Du J,Fan J,et al.The effect of proinflammatory cytokines on IL-17RA expression in NSCLC[J].Med Oncol,2014,31(9):144.
[7] Jiang YX,Li PA,Yang SW,et al.Increased chemokine receptor IL-17RA expression is associated with poor survival in gastric cancer patients[J].Int J Clin Exp Pathol,2015,8(6):7002-7008.
[8] Liu Y,Zhao X,Sun X,et al.Expression of IL-17A,E,and F and their receptors in human prostatic cancer:Comparison with benign prostatic hyperplasia[J].Prostate,2015,75(16):1844-1856.
[9] Wang K,Kim MK,Di CG,et al.Interleukin-17 receptor a signaling in transformed enterocytes promotes early colorectal tumorigenesis[J].Immunity,2014,41(6):1052-1063.
[10] 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.
[11] Asukai K,Kawamoto K,Eguchi H,et al.Prognostic impact of peritumoral IL-17-positive cells and IL-17 axis in patients with intrahepatic cholangiocarcinoma[J].Ann Surg Oncol,2015,22(Suppl 3):S1524-S1531.
[12] Fabre J,Giustiniani J,Garbar C,et al.Targeting the tumor microenvironment:The protumor effects of IL-17 related to cancer type[J].Int J Mol Sci,2016,17(9):1433.
[13] Dong C.TH17 cells in development:An updated view of their molecular identity and genetic programming[J].Nat Rev Immunol,2008,8(5):337-348.
[14] Zhang GQ,Han F,Fang XZ,et al.CD4+,IL17 and Foxp3 expression in different pTNM stages of operable non-small cell lung cancer and effects on disease prognosis[J].Asian Pac J Cancer Prev,2012,13(8):3955-3960.
[15] Zhao SY,He D,Sun W.High expression of L-17RA with poor prognosis of non-small-cell lung cancer[J].Journal of Xinjiang Medical University,2016,9:1151-1154,1158.[赵书源,何丹,孙伟.白细胞介素17受体A在非小细胞肺癌中的表达及其与预后的关系[J].新疆医科大学学报,2016,9:1151-1154,1158.]
[16] Li J,Lau GK,Chen L,et al.Interleukin 17A promotes hepatocellular carcinoma metastasis via NF-kB induced matrix metalloproteinases 2 and 9 expression[J].PLoS One,2011,6(7):e21816.
[17] Wang ZD,Huang C,Li ZF,et al.Chrysanthemum indicum ethanolic extract inhibits invasion of hepatocellular carcinoma via regulation of MMP/TIMP balance as therapeutic target[J].Oncol Rep,2010,23(2):413-421.
[18] Zhang LC,Adili·S,Zhang GQ,et al.Expression and clinical significance of IL-17 and MMP-9 in NSCLC patients of Han and Uighur[J].Cancer Recearch on Prevention and Treatment,2012,7:798-801.[张鲁昌,阿迪力·萨来,张国庆,等.IL-17、MMP-9在汉族和维吾尔族非小细胞肺癌患者中的表达及临床意义[J].肿瘤防治研究,2012,7:798-801.]