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目的探讨Toll样受体5(TLR5)对间充质干细胞(MSC)生物学特性的影响,以期探究TLR5在MSC与肿瘤关系中的作用。方法构建过表达TLR5的慢病毒载体并用相关病毒感染MSC,通过荧光表达量、RT-q PCR和Western印迹验证TLR5在MSC中的过表达情况,通过MSC-TLR5增殖、表面抗原和细胞分化来验证TLR5过表达对MSC生物学性状影响,通过检测CBLB502激活TLR5后白细胞介素6(IL-6)和IL-8的表达水平验证TLR5生物学功能。结果 TLR5慢病毒感染MSC后,mRNA水平和蛋白水平检测TLR5表达量均显著升高,MSC-TLR5增殖、免疫表型和分化能力未发生改变,同时,CBLB502激活TLR5后,IL-6和IL-8表达量显著升高(P<0.001)。结论携带TLR5的慢病毒感染MSC后不影响MSC生物学功能,TLR5具有功能活性且可激活下游信号通路发挥免疫调节作用。
Objective To investigate the effect of Toll-like receptor 5 (TLR5) on the biological characteristics of mesenchymal stem cells (MSCs) in order to explore the role of TLR5 in the relationship between MSC and tumor. Methods The TLR5-overexpressing lentiviral vector was constructed and infected with MSC. The overexpression of TLR5 in MSC was confirmed by fluorescence quantitative analysis, RT-qPCR and Western blotting, and the results were verified by MSC-TLR5 proliferation, surface antigen and cell differentiation TLR5 overexpression on the biological characteristics of MSC, CBLB502 activated TLR5 by detecting the expression of interleukin 6 (IL-6) and IL-8 to verify the biological function of TLR5. Results After TLR5 lentivirus infected MSCs, the mRNA and protein levels of TLR5 were significantly increased, while the proliferation, immunophenotype and differentiation of MSC-TLR5 did not change. TLR5 activated IL-6 and IL- 8 expression was significantly increased (P <0.001). Conclusion TLR5-infected lentivirus does not affect the biological function of MSC. TLR5 has the function of activation and can activate the downstream signaling pathway to play an immunomodulatory role.