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目的:探讨人甲状腺乳头状癌(PTC)细胞中TLR4信号通路对Foxp3表达的调控作用。方法:以人甲状腺乳头状癌K1细胞株为实验材料,选用脂多糖(LPS)作为配体来激活TLR4信号通路,采用RT-PCR方法检测LPS在不同浓度(0,5,10,20,40μg/mL)和不同时间点(12,24,36,48 h)Foxp3的mRNA表达量,流式细胞术检测相应浓度和时间点的Foxp3蛋白和TLR4蛋白表达量的变化;加入LPS抑制剂多粘菌素B(PMB)后Foxp3和mRNA和蛋白的表达量分别利用RT-PCR和流式细胞术检测。结果:10μg/mL的LPS可以显著上调PTC细胞中Foxp3 mRNA和蛋白的表达量(P<0.05),在第24小时达到最佳;LPS作用下,TLR4表达量上调;PMB阻断TLR4信号通路后,Foxp3蛋白表达量较未阻断组显著降低(P<0.05)。结论:在甲状腺乳头状癌K1细胞株中,TLR4作为上游信号调控因子,通过自身表达量改变,进而参与调控Foxp3的表达。
AIM: To investigate the regulatory effect of TLR4 signaling pathway on Foxp3 expression in human thyroid papillary carcinoma (PTC) cells. Methods: Human papillary thyroid carcinoma K1 cell line was used as experimental material, lipopolysaccharide (LPS) was used as ligand to activate TLR4 signaling pathway, and RT-PCR was used to detect the expression of LPS in different concentrations (0, 5, 10, 20, 40μg / mL) and Foxp3 mRNA at different time points (12,24,36,48 h). The expressions of Foxp3 protein and TLR4 protein at the corresponding concentration and time point were detected by flow cytometry. Foxp3 and mRNA and protein expression levels after PMB were detected by RT-PCR and flow cytometry, respectively. Results: LPS at 10μg / mL significantly up-regulated the expression of Foxp3 mRNA and protein in PTC cells (P <0.05), and reached the peak at the 24th hour. The expression of TLR4 was up-regulated after LPS treatment. PMB blocked the TLR4 signaling pathway , Foxp3 protein expression was significantly lower than the non-blocking group (P <0.05). Conclusion: In thyroid papillary carcinoma K1 cell line, TLR4 acts as an upstream signal regulator, which may be involved in the regulation of Foxp3 through its own expression.