肿瘤坏死因子-α对乳腺癌细胞CD44表达调控及其影响细胞迁徙机制的探讨  

Regulation of tumor necrosis factor-α on CD44 molecules expression and impaction on breast cancer cell migration

作　　者：李俊[1] 缪苏宇[2] 陈薇[1] 束永前[1] 王水[2] 殷咏梅[1] 

机构地区：[1]南京医科大学第一附属医院肿瘤科,江苏南京210029 [2]南京医科大学第一附属医院乳腺外科,江苏南京210029

出　　处：《中华肿瘤防治杂志》2010年第8期566-570,共5页Chinese Journal of Cancer Prevention and Treatment

基　　金：国家自然科学基金面上项目[30772474(BA07)];江苏省科技厅基础研究计划(自然科学基金BK2008477);人事部留学回国人员基金2009(IA09);江苏省卫生厅开放课题(XK18200904)

摘　　要：目的:检测肿瘤坏死因子(TNF-α)对乳腺癌细胞株MDA-MB-231中CD44s、CD44V3和CD44V6表达的调控及其对细胞迁徙力的影响,并探讨TNF-α调节CD44可能存在的信号途径。方法:用RT-PCR和蛋白质印迹法检测TNF-α(0、2、20和200ng/mL)作用乳腺癌细胞后其CD44mRNA和蛋白的表达情况,Transwell方法检测TNF-α作用后细胞迁徙能力的改变;以浓度为20ng/mL的TNF-α处理MCF-7和MDA-MB-231后,用蛋白质印迹法检测MAPK(JNK、p38和ERK)信号通路中蛋白磷酸化水平变化,对磷酸化水平增加的蛋白用特异性抑制剂预处理后检测CD44的表达及细胞迁徙能力的改变。结果:TNF-α作用后MDA-MB-231细胞CD44s、CD44V3、CD44V6mRNA和蛋白表达水平均上调;迁徙力增加了22%～90%;TNF-α作用MDA-MB-231细胞后p38磷酸化水平升高;用p38的特异性抑制剂预处理后可以逆转CD44表达和细胞迁徙力的改变。结论:TNF-α通过p38途径对MDA-MB-231细胞CD44表达进行调控,CD44的表达可以增强肿瘤细胞的迁徙力。OBJECTIVE：To detect CD44s,CD44V3 and CD44V6 expression in breast cancer cell line MDA-MB-231 treated with TNF-α and the effect on cell migration,and investigate the possible pathways via which TNF-α induces CD44 expression in MDA-MB-231 cell.METHODS：CD44 mRNA was detected by RT-PCR,and CD44 protein expression was detected by Western blot assay after treated with TNF-α （0,2,20 and 200 ng/mL）.Transwell was used for detecting the migration of cells in vitro.MDA-MB-231 cell was treated with TNF-α （20 ng/mL） for 24 h.Western blot was used for detecting the phosphorylation of MAPK pathway proteins （JNK,p38,ERK）.The CD44 expression and the migration of cells in vitro were detected after pre-treatment with the specific inhibitors of activated phosphorylation protein.RESULTS：TNF-α treatment increased the levels of CD44s,CD44V3 and CD44V6 mRNAs and proteins in MDA-MB-231 cells.The migration of MDA-MB-231 cells increased by 22%-90%.When treated with TNF-α,p38 phosphorylation of MDA-MB-231 cells increased.Both of CD44 expression and cell migration induced by TNF-α were reduced after the pre-treatment with paired specific inhibitors.CONCLUSIONS：TNF-α regulates the CD44 expression via p38 pathway in MDA-MB-231 cell.CD44 premotes BC cells migration.

关 键 词：乳腺肿瘤/病理学 肿瘤坏死因子Α 抗原 CD44 肿瘤细胞 培养的 

分 类 号：R737.9[医药卫生—肿瘤]