肿瘤细胞来源外泌体miR-21诱导巨噬细胞焦亡促进胃癌细胞转移机制的实验研究OA
Experimental Study on the Mechanism of Tumor Cell-Derived Exosomes miR-21 Inducing Macrophage Pyroptosis to Promote Metastasis in Gastric Cancer Cells
目的 探究肿瘤细胞来源外泌体miR-21在胃癌细胞转移过程中对巨噬细胞焦亡的影响.方法 培养人胃癌细胞HGC-27和人正常胃黏膜上皮细胞GES-1,提取两种细胞来源外泌体,通过qRT-PCR检测外泌体内miR-21表达水平.通过慢病毒转染细胞的方法构建稳定过表达miR-21的HGC-27细胞株及阴性对照细胞株,之后提取外泌体,并使用PKH67染料标记外泌体.根据来源细胞株不同,将外泌体分为miR-21-exo组、miR-21-mimics-exo组和miR-21-mimicsNC-exo组.使用佛波脂(PMA)诱导THP-1细胞分化为M0型巨噬细胞,并将M0型巨噬细胞分为exo-miR-21组、exo-miR-21-mimics组、exo-miR-21-mimicsNC组.加入外泌体干预处理36 h后,通过Western blot和ELISA检测巨噬细胞焦亡相关因子的表达水平.提取上述各组巨噬细胞培养上清,与HGC-27细胞共培养,划痕试验和Transwell试验检测HGC-27细胞迁移和侵袭能力.结果 与GES-1细胞来源外泌体相比,miR-21在HGC-27细胞来源外泌体内高表达,差异具有统计学意义(t=12.00,P<0.001).与exo-miR-21组和exo-miR-21-mimicsNC组相比,exo-miR-21-mimics组巨噬细胞焦亡相关因子表达水平显著升高,差异具有统计学意义(t=19.23～26.79,均P<0.000 1).与添加了exo-miR-21组和exo-miR-21-mimicsNC组巨噬细胞培养上清的HGC-27细胞相比,添加了exo-miR-21-mimics组巨噬细胞培养上清的HGC-27细胞迁移和侵袭能力显著增强,差异具有统计学意义(t迁移=11.39、11.16,t侵袭=10.80、9.153,均P<0.001).结论 肿瘤细胞来源外泌体miR-21能够通过诱导巨噬细胞焦亡来促进胃癌细胞转移.
Objective To explore the effect of tumor cell-derived exosomal miR-21 on macrophage pyroptosis during gastric can-cer cell metastasis.Methods Human gastric cancer cell line HGC-27 and human normal gastric epithelial cell GES-1 were cul-tured and exosomes were extracted.The expression of miR-21 in exosomes was detected by qRT-PCR.HGC-27 cell line stably over expressing miR-21 and negative control cell line were constructed.Exosomes were extracted and labeled with PKH67 dye.The exosomes were divided into miR-21-exo group,miR-21-mimics-exo group and miR-21-mimicsNC-exo group.THP-1 cells were induced to differentiate into M0 macrophages using PMA and grouped into exo-miR-21,exo-miR-21-mimics,exo-miR-21-mimicsNC.After 36 hours of treatment with exosomes,the expression levels of macrophage scorch-related factors were detected by Western blot and ELISA.The supernatants were extracted and co-cultured with HGC-27 cells.The migration and invasion of HGC-27 cells were detected by scratch test and Transwell test.Results MiR-21 was highly expressed in HGC-27 cell-derived exosomes compared with GES-1 cell-derived exosomes,and the difference was statistically significant(t=12.00,P<0.001).Com-pared with the exo-miR-21 group and the exo-miR-21-mimicsNC group,the expression level of macrophage pyroptosis-related factors in the exo-miR-21-mimics group was significantly higher,the differences were statistically significant(t=19.23～26.79,all P<0.000 1).Compared with HGC-27 cells added with macrophage culture supernatants from the exo-miR-21 intervention group and the exo-miR-21-mimicsNC intervention group,HGC-27 cells with the addition of exo-miR-21-mimics had significantly en-hanced migration and invasion ability in the supernatant of macrophage culture,the differences were statistically significant(tmigra-tion=11.39,11.16,tinvasion=10.80,9.153,all P<0.001).Conclusions Tumor cell-derived exosomal miR-21 are able to promote gastric cancer cell metastasis by inducing macrophage pyroptosis.
张伟;蔡振花;杨晨;李浩闯;魏琦;王玉宏
邯郸市中心医院 普外七科,河北 邯郸 056001邯郸市中心医院 护理部,河北 邯郸 056001邯郸市中心医院 肿瘤二科,河北 邯郸 056001邯郸市中心医院 普外七科,河北 邯郸 056001邯郸市人民医院普外科,河北 邯郸 056001邯郸市中心医院 普外七科,河北 邯郸 056001
医药卫生
外泌体微小RNA-21巨噬细胞细胞焦亡胃癌
exosomesmiR-21macrophagespyroptosisgastric cancer
《现代检验医学杂志》 2026 (1)
29-34,6
河北省卫生健康委2022年度医学课题(20220017).
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