AXL对乳头状甲状腺癌STAT3信号通路活化的促进作用
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摘要
目的探讨AXL对乳头状甲状腺癌(papillary thyroid cancer,PTC) STAT3表达的影响。方法采用基因集富集分析的方法(gene set enrichment analysis,GSEA)对PTC转录组数据库中的572个PTC样本AXL的表达进行比较研究,分析AXL对PTC STAT3表达的影响。HEK293T细胞随机分为空白对照组、空载体组和pc DNA-AXL组(AXL组),Western blot检测各组p-STAT3的表达;用不同浓度的AXL配体GAS6处理BCPAP细胞系,Western blot检测细胞p-STAT3和核内STAT3的表达。结果 GSEA分析发现AXL高表达的PTC其STAT3信号通路活化程度更高(normalized enrichment score,NES)=1. 47,P <0. 05)。HEK293T细胞转染AXL后,AXL高表达导致p-STAT3活性增高(P <0. 05)。AXL配体GAS6处理BCPAP细胞系后,p-STAT3活性显著增高(P <0. 05),STAT3核转位增加(P <0. 01)。结论 AXL可促进PTCSTAT3信号通路的活化。
Objective Signal transducer and activator of transcription( STAT3) signaling is hyperactivated in papillary thyroid cancer(PTC),but the mechanism of which is not fully understood. AXL is upregualted in PTC which activates several signaling pathways. The objective of this article is to investigate whether AXL activates STAT3 signaling pathway. Methods The AXL of 572 PTC samples of the transcriptomes of PTC was analyzed by gene set enrichment analysis(GSEA)to assess the effect of AXL on activation of STAT3 signaling pathway. HEK293 T cells were transfected by AXL plasmid and p-STAT3 level was determined by Western blot. PTC cell line BCPAP was treated with GAS6,and p-STAT3 and nuclear STAT3 were determined by Western blot. Results GSEA analysis found STAT3 activation was higher in AXL-high PTCs(Normalized enrichment score(NES) = 1. 47,P < 0. 05). HEK293 T cells transfected with AXL displayed higher level of p-STAT3(P < 0. 05). Furthermore,treatment of PTC cell line BCPAP with GAS6,the ligand of AXL,led to increased pSTAT3(P < 0. 05) and more STAT3 nuclear translocation(P < 0. 05). Conclusion AXL increased STAT3 activation in PTC.
Objective Signal transducer and activator of transcription( STAT3) signaling is hyperactivated in papillary thyroid cancer(PTC),but the mechanism of which is not fully understood. AXL is upregualted in PTC which activates several signaling pathways. The objective of this article is to investigate whether AXL activates STAT3 signaling pathway. Methods The AXL of 572 PTC samples of the transcriptomes of PTC was analyzed by gene set enrichment analysis(GSEA)to assess the effect of AXL on activation of STAT3 signaling pathway. HEK293 T cells were transfected by AXL plasmid and p-STAT3 level was determined by Western blot. PTC cell line BCPAP was treated with GAS6,and p-STAT3 and nuclear STAT3 were determined by Western blot. Results GSEA analysis found STAT3 activation was higher in AXL-high PTCs(Normalized enrichment score(NES) = 1. 47,P < 0. 05). HEK293 T cells transfected with AXL displayed higher level of p-STAT3(P < 0. 05). Furthermore,treatment of PTC cell line BCPAP with GAS6,the ligand of AXL,led to increased pSTAT3(P < 0. 05) and more STAT3 nuclear translocation(P < 0. 05). Conclusion AXL increased STAT3 activation in PTC.
引文
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[23]Scaltriti M,Elkabets M,Baselga J.Molecular pathways:AXL,a membrane receptor mediator of resistance to therapy[J].Clin Cancer Res,2016,22(6):1313-7.DOI:10.1158/1078-0432.CCR-15-1458.
[24]Bivona T,Okimoto R.AXL receptor tyrosine kinase as a therapeutic target in NSCLC[J].Lung Cancer:Targets and Therapy,2015,6:27-34.DOI:10.2147/LCTT.S60438.
[25]Wu X,Liu X,Koul S,et al.AXL kinase as a novel target for cancer therapy[J].Oncotarget,2014,5(20):9546-9563.DOI:10.18632/oncotarget.2542.
[2]代丽丽,张国安,崔文.BRAFV600E和TERT启动子突变与甲状腺乳头状癌的关系[J].济宁医学院学报,2018,41(4):290-293.DOI:10.3969/j.issn.1000.9760.2018.04.016.
[3]Chen W,Zheng R,Baade PD,et al.Cancer statistics in China,2015[J].CA Cancer J Clin,2016,66(2):115-132.DOI:10.3322/caac.21338.
[4]Xing M.BRAF mutation in thyroid cancer[J].Endocr Relat Cancer,2005,12(2):245-262.DOI:10.1677/erc.1.0978.
[5]Hafizi S,Dahlback B.Signalling and functional diversity within the Axl subfamily of receptor tyrosine kinases[J].Cytokine Growth Factor Rev,2006,17(4):295-304.DOI:10.1016/j.cytogfr.2006.04.004.
[6]Paccez JD,Vogelsang M,Parker MI,et al.The receptor tyrosine kinase Axl in cancer:biological functions and therapeutic implications[J].Int J Cancer,2014,134(5):1024-1033.DOI:10.1002/ijc.28246.
[7]Graham DK,Deryckere D,Davies KD,et al.The TAMfamily:phosphatidylserine sensing receptor tyrosine kinases gone awry in cancer[J].Nat Rev Cancer,2014,14(12):769-85.DOI:10.1038/nrc3847.
[8]Qu X,Liu G,Zhong X,et al.Role of AXL expression in non-small cell lung cancer[J].Oncol Lett,2016,12(6):5085-5091.DOI:10.3892/ol.2016.5356.
[9]Zhang Z,Lee JC,Lin L,et al.Activation of the AXL kinase causes resistance to EGFR-targeted therapy in lung cancer[J].Nat Genet,2012,44(8):852-860.DOI:10.1038/ng.2330.
[10]Yu H,Lee H,Herrmann A,et al.Revisiting STAT3 signalling in cancer:new and unexpected biological functions[J].Nat Rev Cancer,2014,14(11):736-746.DOI:10.1038/nrc3818.
[11]Trovato M,Grosso M,Vitarelli E,et al.Distinctive expression of STAT3 in papillary thyroid carcinomas and a subset of follicular adenomas[J].Histol Histopathol,2003,18(2):393-399.DOI:10.14670/HH-18.393.
[12]王翡,罗大虎,刘雪梅.甲状腺乳头状癌组织中STAT3和核因子κB水平增加[J].细胞与分子免疫学杂志,2016,32(10):1386-1389.
[13]Subramanian A,Tamayo P,Mootha VK,et al.Gene set enrichment analysis:a knowledge-based approach for interpreting genome-wide expression profiles[J].Proc Natl Acad Sci USA,2005,102(43):15545-15550.DOI:10.1073/pnas.0506580102.
[14]Zhang G,Kong X,Wang M,et al.AXL is a marker for epithelial-mesenchymal transition in esophageal squamous cell carcinoma[J]Oncol Lett,2018,15(2):1900-1906.DOI:10.3892/ol.2017.7443.
[15]Agrawal N,Akbani R,Aksoy BA,et al.Integrated genomic characterization of papillary thyroid carcinoma[J].Cell,2014,159(3):676-690.DOI:10.1016/j.cell.2014.09.050.
[16]冯春琼,邹亚光,周其赵,等.GSEA在全基因组表达谱芯片数据分析中的应用[J].现代生物医学进展,2009,9(13):2553-2557.
[17]Avilla E,Guarino V,Visciano C,et al.Activation of TYRO3/AXL tyrosine kinase receptors in thyroid cancer[J].Cancer Res,2011,71(5):1792-1804.DOI:10.1158/0008-5472.CAN-10-2186.
[18]Tanaka K,Nagayama Y,Nakano T,et al.Expression profile of receptor-type protein tyrosine kinase genes in the human thyroid[J].Endocrinology,1998,139(3):852-858.DOI:10.1210/endo.139.3.5791.
[19]Isildak SM,Tutuncu NB,Altundag O.Tyrosine kinase inhibitors in thyroid cancer:may Axl/Gas6 pathway be a hidden target[J].UHOD-ULUSLARARASI HEMATOLOJI-ONKOLOJI DERGISI,2013,23(3):207-212.
[20]Zhang G,Wang M,Zhao H,et al.Function of Axl receptor tyrosine kinase in non-small cell lung cancer[J].Oncol Lett,2018,15(3):2726-2734.DOI:10.3892/ol.2017.7694.
[21]Gay CM,Balaji K,Byers LA.Giving AXL the axe:targeting AXL in human malignancy[J].Br J Cancer,2017,116:415.DOI:10.1038/bjc.2016.428.
[22]Antony J,Huang RY.AXL-driven EMT state as a targetable conduit in cancer[J].Cancer Res,2017,77(14):3725-3732.DOI:10.1158/0008-5472.CAN-17-0392.
[23]Scaltriti M,Elkabets M,Baselga J.Molecular pathways:AXL,a membrane receptor mediator of resistance to therapy[J].Clin Cancer Res,2016,22(6):1313-7.DOI:10.1158/1078-0432.CCR-15-1458.
[24]Bivona T,Okimoto R.AXL receptor tyrosine kinase as a therapeutic target in NSCLC[J].Lung Cancer:Targets and Therapy,2015,6:27-34.DOI:10.2147/LCTT.S60438.
[25]Wu X,Liu X,Koul S,et al.AXL kinase as a novel target for cancer therapy[J].Oncotarget,2014,5(20):9546-9563.DOI:10.18632/oncotarget.2542.