PPAR-γ抑制高糖微环境下NCI-H460细胞增殖的分子机制

PPAR-γinhibits the proliferation of human large cell lung cancer NCI-H460 cells through the NLRP3 inflammasome in a high-glucose microenvironment

目的探讨PPAR-γ在高糖微环境下对人大细胞肺癌NCI-H460细胞增殖的影响及其分子机制。方法分别用正常糖(空白组)、高渗(对照组)和30 mmol/L葡萄糖(高糖组)培养基处理NCI-H460细胞,用CCK-8法和平板克隆实验分析高糖微环境对NCI-H460细胞增殖能力的影响;用Western blotting检测NLRP3炎症小体相关蛋白和PPAR-γ蛋白的表达;用特异性PPAR-γ激活剂罗格列酮刺激高糖微环境下的NCI-H460细胞;用Western blotting检测NLRP3炎症小体相关蛋白的表达,CCK-8法和平板克隆实验分析高糖微环境下PPAR-γ对NCI-H460细胞增殖能力的影响;用NLRP3炎症小体激动剂NSS联合罗格列酮刺激高糖微环境下的NCI-H460细胞,CCK-8法和平板克隆实验分析NLRP3炎症小体是否参与高糖微环境下PPAR-γ对NCI-H460细胞增殖的抑制作用。结果高糖微环境能够显著提高NCI-H460细胞的增殖能力,诱导NLRP3炎症小体活性升高,下调PPAR-γ蛋白表达,PPAR-γ激活剂罗格列酮能有效抑制NLRP3炎症小体的活性,抑制NCI-H460细胞的增殖,且这一作用可被NLRP3炎症小体激动剂逆转。结论PPAR-γ通过下调NLRP3炎症小体活性,抑制高糖微环境下人大细胞肺癌NCI-H460细胞增殖。

Objective To investigate the effect of PPAR-γon the proliferation of human large cell lung cancer(NCI-H460)cells in a high-glucose microenvironment and to explore the associated molecular mechanism.Methods NCI-H460 cells were treated with normal-glucose(blank group),hypertonic(control group),and high-glucose(30 mmol/L;high-glucose group)media.The effects of a high-glucose microenvironment on the proliferation of NCI-H460 cells were analyzed using Cell Counting Kit 8(CCK-8)and colony-formation assays.The specific PPAR-γactivator,rosiglitazone,was used to treat NCI-H460 cells in a high-glucose microenvironment,and the expression of NLRP3-inflammasome-related proteins was detected by Western blotting.The effect of PPAR-γon the proliferation of NCIH460 cells in a high-glucose microenvironment was analyzed by CCK-8 and clony-formation assays.The NLRP3 inflammasome agonist,NSS,combined with rosiglitazone,were used to treat NCI-H460 cells in a high-glucose microenvironment.CCK-8 and clony-formation assays were used to analyze whether the NLRP3 inflammasome was involved in the inhibitory effect of PPAR-γon the proliferation of NCIH460 cells in a high-glucose microenvironment.Results A high-glucose microenvironment significantly induced the proliferation of NCI-H460 cells,increased the activity of the NLRP3 inflammasome,and reduced the expression levels of PPAR-γprotein.Rosiglitazone effectively inhibited NLRP3 inflammasome activity and NCI-H460 cell proliferation,and this effect was reversed by NLRP3 inflammasome agonist.Conclusion PPAR-γinhibits the proliferation of NCI-H460 cells in a high-glucose microenvironment by down-regulating the activity of the NLRP3 inflammasome.