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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 investigate the effects of tumor necrosis factor (TNF-α) on the expression of CD44s, CD44V3 and CD44V6 in breast cancer cell line MDA-MB-231 and its effect on cell migration. way. Methods: The expression of CD44 mRNA and protein in breast cancer cells treated with TNF-α (0, 2, 20 and 200 ng / mL) was detected by RT-PCR and Western blotting. Transwell method was used to detect the migration ability of cells The protein phosphorylation levels of MAPK (JNK, p38 and ERK) in MAPK (JNK, p38 and ERK) signaling pathways were detected by Western blotting, and the phosphorylation level The increased protein was pretreated with specific inhibitors to detect CD44 expression and cell migration ability changes. Results: The mRNA and protein expressions of CD44s, CD44V3 and CD44V6 in MDA-MB-231 cells were upregulated after TNF-α treatment. Migration increased by 22% -90%. The level of p38 phosphorylation Increased; p38 specific inhibitor pretreatment can reverse the CD44 expression and cell migration changes. Conclusion: TNF-α regulates CD44 expression in MDA-MB-231 cells via p38 pathway. The expression of CD44 enhances the migration ability of tumor cells.