丙泊酚作用NLRP3/ASC/caspase-1通路对肺癌A549细胞焦亡的影响OACSTPCD
Effect of propofol on pyroptosis of lung cancer A549 cells by NLRP3/ASC/caspase-1 pathway
目的 探讨丙泊酚作用核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)/凋亡相关斑点样蛋白(ASC)/半胱氨酸天冬氨酸蛋白酶-1(caspase-1)通路对A549细胞焦亡的影响.方法 超低吸附培养法建立肺癌A549细胞三维培养模型,建立模型后采用不同浓度丙泊酚作用肺癌A549细胞,CCK-8法检测丙泊酚对A549细胞增殖的影响;酶联免疫吸附法检测肺癌A549细胞上清炎性细胞因子白细胞介素(IL)-18、IL-1β、IL-6表达水平变化;蛋白免疫印迹法检测各组焦亡相关蛋白NLRP3、ASC、caspase-1、焦孔素蛋白N端(GSDMD-N)、IL-1β表达水平.结果 三维培养下,A549细胞间有聚拢黏附趋势,培养12 h即可清晰观察到细胞球.与空白对照组相比,随着丙泊酚浓度增加,肺癌A549细胞存活率依次降低(P<0.05);肺癌A549细胞上清炎性细胞因子 IL-18、IL-1β、IL-6、焦亡相关蛋白NLRP3、ASC、caspase-1、GSDMD-N、IL-1β表达水平依次增加(P<0.05).结论 超低吸附培养法成功建立肺癌A549细胞三维培养模型,丙泊酚通过激活NLRP3/ASC/caspase-1通路促进细胞焦亡,进而抑制肺癌A549细胞增殖.
Objective To investigate the effect of propofol on pyroptosis and A549 cells via the NLRP3/ASC/caspase-1 pathway.Methods Establish a three-dimensional culture model of A549 tumor cells using ultra-low attachment plates,A549 cells were cultured using ultra-low adsorption culture plates to establish a three-dimensional culture model.The CCK-8 method was used to detect the effect of propofol on A549 cell proliferation;the inflammatory factors interleukin(IL)-18,IL-1β,and IL-6 were detected in the A549 lung cancer cell supernatants using enzyme-linked immunosorbent assays;western blotting was used to detect the expression levels of pyrolysis-asso-ciated proteins NLRP3,ASC,caspase-1,GSDMD-N,and IL-1β in A549 lung cancer cells in each group.Results Compared to the blank control group,the survival rate of A549 cells in low,medium,and high concentrations of propofol in each group decreased in turn(P<0.05);the levels of inflammatory factors IL-18,IL-1β,and IL-6 in the A549 cell supernatant,and scorch related protein NLRP3,ASC,caspase-1,GSDMD-N,and IL-1β increased with the increased propofol concentrations(P<0.05).Conclusion The three-dimensional culture model of lung cancer A549 cells was successfully established using the ultra-low adsorption culture method.Propofol can promote cell apoptosis and inhibit the pyroptosis of A549 lung cancer cells via activating the NLRP3/ASC/caspase-1 pathway.
关冀弛;杨金雨;刘丹;李磊;李东华;汪海峰
沈阳化工大学制药与生物工程学院,沈阳 110142
临床医学
丙泊酚超低吸附培养法肿瘤细胞三维培养模型肺癌细胞焦亡
propofolultra-low attachment platesthree-dimensional culture model of tumor cellslung cancerpyroptosis
《中国医科大学学报》 2024 (002)
132-135,141 / 5
辽宁省教育厅科学研究经费项目青年科技人才"育苗"项目(LQ2020021)
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