顺铂诱导铁死亡促进肿瘤相关巨噬细胞极化抑制宫颈癌细胞耐药性

目的:探究顺铂通过引起宫颈癌细胞铁死亡进而诱导肿瘤相关巨噬细胞极化从而抑制宫颈癌细胞耐药性的机制。方法:使用5μmol/L顺铂处理顺铂耐药宫颈癌细胞Hela/R一定时间后,采用qRT-PCR法检测铁死亡相关基因的mRNA表达变化情况;使用ELISA试剂盒和荧光染色法测定铁死亡后的Hela/R细胞释放高迁移率族蛋白1(high mobility group box 1,HMGB1)的情况。将顺铂处理过的Hela/R细胞和M2型小鼠骨髓来源巨噬细胞共培养一定时间后,采用流式细胞术检测巨噬细胞激活情况。将共培养后的巨噬细胞和Hela/R细胞共孵育,采用CCK8法和流式细胞术分别检测肿瘤细胞的存活率和凋亡情况。结果:实验数据显示,顺铂可引起Hela/R细胞的铁死亡,抑制其铁死亡抑制基因Slc40a1、Slc7a11、Slc3a2、Gpx4、Fth1、Blvrb的mRNA表达(P<0.05),上调铁死亡诱发基因Slc5a1、Tfrc的mRNA表达(P<0.01)。铁死亡Hela/R细胞释放损伤相关模式分子HMGB1,诱导M2型肿瘤相关巨噬细胞的CD80、CD86和CD40平均荧光强度提升,增强了肿瘤相关巨噬细胞对Hela/R细胞的杀伤能力。结论:顺铂通过引起宫颈癌细胞的铁死亡激活肿瘤相关巨噬细胞进而达到有效杀伤肿瘤细胞的效果。

Cisplatin-induced ferroptosis promotes tumor-associated macrophage polarization to inhibits drug resistance in cervical cancer cells

Objective:Explore the mechanism by which cisplatin inhibits cervical cancer by means of causing the ferroptosis of cervical cancer cells to induce the polarization of tumor-associated macrophages.Methods:Cisplatin-resistant cervical cancer cells Hela/R were treated with 5 μmol/L cisplatin for a certain period of time,qRT-PCR was used to detect the mRNA expression changes of iron death-related genes.ELISA kit and fluorescence staining method were used to determine the release of HMGB1 from Hela/R cells after ferroptosis.Cisplatin-treated Hela/R cells and M2 mouse bone marrow-derived macrophages were co-cultured,and the activation of macrophages were detected by flow cytometry.The educated macrophages and Hela/R cells were co-cultured,and the survival rate and apoptosis of tumor cells were detected by CCK8 method and flow cytometry,respectively.Results:Experimental data shows that cisplatin cause ferroptosis in Hela/R cells,inhibiting the mRNA expression of the iron death suppressor genes Slc40a1,Slc7a11,Slc3a2,Gpx4,Fth1 and Blvrb（P＜0.05),and up-regulating the iron death-inducing genes Slc5a1,Tfrc mRNA expression（P＜0.01).Hela/R which undertook ferroptosis released the damage-related pattern molecule HMGB1,which induces an increase of mean fluorescence intensity of CD80,CD86 and CD40 in M2-like tumor-related macrophages,resulting in enhanced antitumor ability of tumor-related macrophages to Hela/R cells.Conclusion:Cisplatin can activate tumor-associated macrophages by inducing iron death of cervical cancer cells to achieve effective tumor cell killing effect.