摘要
该文探讨肿瘤坏死因子α(TNFα)活化信号转导和转录激活因子3(STAT3)的分子机制。采用流式细胞术(FACS)检测TNF受体TNFR1在鼻咽癌细胞5-8F和宫颈癌细胞HeLa中的蛋白表达水平;qRT-PCR检测TNFα对其受体TNFR1和TNFR2的mRNA水平的影响;ELISA检测细胞因子白细胞介素8(IL-8)的蛋白水平;Western blot检测受体和信号转导分子的总蛋白水平及蛋白磷酸化水平。结果显示,5-8F和HeLa细胞表达功能性的TNF受体和表皮生长因子受体(EGFR);TNFα处理细胞可诱导STAT3的活化,且呈时间和剂量依赖性;TNFα也能活化EGFR,用EGFR的抑制剂进行处理,逆转了TNFα诱导的EGFR(Y1068)的磷酸化,也逆转了STAT3的磷酸化;进一步研究结果显示,TNFα可活化促癌酪氨酸蛋白激酶SRC,用SRC抑制剂处理,逆转了TNFα诱导的EGFR活化及其下游STAT3的磷酸化。总之,在肿瘤细胞中存在TNFα-SRC-EGFR-STAT3信号转导通路,提示EGFR可能是炎症诱导肿瘤的桥梁。
This study explored the molecular mechanism of STAT3 signal transduction induced by TNFα (tumor necrosis factor α).FACS was employed to determine the expression of TNF receptor TNFR1 in 5-8F nasopharyngeal carcinoma cells and HeLa cervical cancer cells.qRT-PCR was used to measure the mRNA levels of TNFR1 and TNFR2 induced by TNFα.ELISA was used to measure the protein levels of cytokine IL-8.Western blot was used to detect the total and phosphorylated protein levels of receptors and signal molecules.The results showed that 5-8F and HeLa cells expressed functional TNF receptor and EGFR.The treatment of cells with TNFα induced the activation of STAT3 in a time-dependent and dose-dependent manner.TNFα could also activate EGFR.Treatment with EGFR inhibitors reversed the phosphorylation of EGFR (Y1068) induced by TNFα and also reversed the phosphorylation of STAT3.Furthermore,TNFα activated the pro-cancer tyrosine protein kinase SRC.After treatment with SRC inhibitors,TNFα-induced EGFR activation and downstream STAT3 phosphorylation were reversed.In conclusion, in certain cancer cells, the signal transduction pathway TNFα-SRC-EGFR-STAT3 is existed, and EGFR may be a bridge for linking inflammation with cancer.
作者
曹婷
张朝
胡玲
蒋斌元
胡锦跃
CAO Ting;ZHANG Zhao;HU Ling;JIANG Binyuan;HU Jinyue(The Department of Clinical Laboratory Affiliated Changsha Central Hospital,Hengyang Medical School,University of South China,Changsha 410004,China;Central Laboratory,Affiliated Changsha Central Hospital,Hengyang Medical School,University of South China,Changsha 410004,China)
出处
《中国细胞生物学学报》
CAS
CSCD
2022年第2期239-247,共9页
Chinese Journal of Cell Biology
基金
国家自然科学基金青年科学基金(批准号:81172042)
湖南省自然科学基金面上项目(批准号:2020JJ4636)资助的课题。
