IL-1β促进肺癌细胞增殖的作用机制研究

发布时间：2018-06-28 11:08

本文选题：巨噬细胞 + 肺癌　；参考：《中国免疫学杂志》2017年01期

【摘要】：目的:研究IL-1β在巨噬细胞促进肺癌细胞增殖的作用机制。方法:采用Transwell屮插入式细胞培养皿共培养人肺癌细胞株A549、NCI-H520细胞和巨噬细胞。Brd U-ELISA法检测巨噬细胞对肺癌细胞的增殖能力。采用Real-time PCR法检测巨噬细胞和A549、NCI-H520细胞中IL-1βmRNA的表达。应用IL-1β的中和抗体抑制IL-1β的活性,观察IL-1β在巨噬细胞促进肺癌细胞增殖中的作用。利用Western blot法检测肺癌细胞中自噬标志物Beclin 1的表达。结果:Brd UELISA检测结果:A549细胞与巨噬细胞以1∶0.5的比例共培养,A549细胞的OD值从(0.41±0.06)增加到(1.13±0.10),差异有统计学意义(P0.05),且A549细胞的OD值随共培养巨噬细胞比例的增加而升高(P0.05)。NCI-H520细胞与不同浓度的巨噬细胞共培养后,NCI-H520细胞的OD值变化趋势与A549细胞相似。Real-time PCR法检测结果:在共培养后,巨噬细胞中IL-1βmRNA的表达显著上调,且有时间依赖性,并显著高于肺癌细胞中的表达。加入IL-1β中和抗体后,Brd U ELISA法检测的细胞增殖,结果显示IL-1β中和抗体可明显抑制巨噬细胞对肺癌细胞A549和NCI-H520的增殖促进作用。Western blot法检测结果显示IL-1β可显著抑制肿瘤细胞Beclin 1的表达。结论:巨噬细胞和肺癌细胞通过增强IL-1β的表达促进肿瘤细胞的增殖。抑制肿瘤细胞的自噬可能是IL-1β促进肺癌发展的作用机制。
[Abstract]:Objective: to study the mechanism of IL-1 尾 on macrophage promoting proliferation of lung cancer cells. Methods: the proliferation of human lung cancer cell line A549 NCI-H520 and macrophage. Brd U-ELISA was used to detect the proliferation of human lung cancer cell line A549 and Brd U-ELISA. Real-time PCR was used to detect the expression of IL-1 尾 mRNA in macrophages and A549 NCI-H 520 cells. The neutralizing antibody of IL-1 尾 was used to inhibit the activity of IL-1 尾 and to observe the role of IL-1 尾 in promoting the proliferation of lung cancer cells. The expression of the autophagy marker Beclin 1 in lung cancer cells was detected by Western blot assay. Results the OD value of A549 cells co-cultured with macrophages was increased from (0.41 卤0.06) to (1.13 卤0.10). The OD value of A549 cells increased with the increase of co-cultured macrophages (P0.05). After co-culture with different concentrations of macrophages, the change trend of OD value of NCI-H520 cells was similar to that of A549 cells detected by .Real-time PCR: after co-culture, the changes of OD value of NCI-H520 cells were similar to those of A549 cells. The expression of IL-1 尾 mRNA in macrophages was significantly up-regulated and time-dependent, and significantly higher than that in lung cancer cells. The cell proliferation was detected by BrdU Elisa after the addition of IL-1 尾 neutralizing antibody. The results showed that IL-1 尾 neutralizing antibody could significantly inhibit the proliferation of lung cancer cells A549 and NCI-H520. Western blot assay showed that IL-1 尾 could significantly inhibit the expression of Beclin1 in tumor cells. Conclusion: macrophages and lung cancer cells promote the proliferation of tumor cells by enhancing the expression of IL-1 尾. Inhibition of autophagy of tumor cells may be the mechanism of IL-1 尾 promoting the development of lung cancer.
【作者单位】：蚌埠医学院生物化学与分子生物学教研室;同济大学附属同济医院检验科;
【基金】：国家自然科学基金(81272603,81472179) 安徽省自然科学基金(KJ2014A162)资助项目
【分类号】：R734.2

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