M1型巨噬细胞外泌体通过JAK1/STAT3信号通路抑制卵巢癌细胞免疫逃逸OA
Study on the Inhibition of Ovarian Cancer Cell Immune Escape by M1 Macrophage Exosomes through JAK1/STAT3 Signaling Pathway
目的 探讨M1型巨噬细胞外泌体调节酪氨酸激酶1(JAK1)/信号转导和转录激活因子3(STAT3)通路对卵巢癌细胞行为的影响.方法 提取M1型巨噬细胞分泌的外泌体(M1-exo),用浓度为0、50、100μg/ml的外泌体与人卵巢癌SKOV3细胞共培养48h,记为PBS组、M1-exo(50μg/ml)组、M1-exo(100μg/ml)组.平板克隆实验检测SKOV3细胞增殖;Transwell实验评估SKOV3细胞的迁移与侵袭特性;流式细胞术分析SKOV3细胞的凋亡状况;Western blot技术检测细胞中p-JAK1、p-STAT3的蛋白表达情况.将自然杀伤细胞NK-92MI分别与上述三组细胞共培养,检测共培养细胞上清白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)水平及共培养体系中NK-92MI的免疫杀伤率.结果 提取的M1-exo展现出典型外泌体的形态特性,并且显著表达了外泌体的标志性蛋白质CD9、CD81和TSG101,同时PKH67标记显示M1-exo可被SKOV3细胞摄取.与PBS组比较,M1-exo(50μg/ml)SKOV3细胞迁移、侵袭及凋亡均被抑制,细胞中p-JAK1、p-STAT3蛋白表达下降(t=4.08～21.63,均P<0.05);与M1-exo(50μg/ml)组比较,M1-exo(100μg/ml)组SKOV3细胞增殖、迁移、侵袭及凋亡均被抑制,细胞中p-JAK1、p-STAT3蛋白表达下降(t=3.61～15.60,均P<0.05).共培养后与PBS组比较,M1-exo(50μg/ml)组、M1-exo(100μg/ml)组上清IL-6、TNF-α水平及NK-92MI细胞免疫杀伤率均增加(t=21.76～14.88,均P<0.05),其中M1-exo(100μg/ml)组上清IL-6、TNF-α水平及NK-92MI细胞免疫杀伤率最大,差异具有统计学意义(t=4.41～6.88,均P<0.05).结论 M1型巨噬细胞外泌体可能通过下调JAK1/STAT3信号通路抑制SKOV3细胞增殖、迁移、侵袭及免疫逃逸,促进细胞凋亡.
Objective To investigate the influence of M1 macrophage-derived exosomes on the JAK1/STAT3 pathway and its impact on ovarian cancer cell behavior.Methods Isolated extracellular vesicles(M1 exo)from M1 macrophages and incubated them with human ovarian cancer SKOV3 cells at 0,50 and 100 μg/ml for 48 hours,labeled as PBS,M1 exo(50 μg/ml)and M1 exo(100 μg/ml)groups.The plate cloning assay was employed to assess SKOV3 cell proliferation,and the Transwell assay for evaluating SKOV3 cell migration and invasion.Flow cytometry was used to measure SKOV3 cell apoptosis,and Western blotting for analyzing p-JAK1 and p-STAT3 protein expression.Additionally,NK-92MI natural killer cells were co-cultured with the aforementioned groups,and the supernatant's IL-6 and TNF-α levels,as well as the immune killing rate of NK-92MI in the co-culture system,were assessed.Results The isolated M1 exo exhibited the typical morphology of extracellular vesicles and strongly expressed marker proteins CD9,CD81 and TSG101.PKH67 staining indicatesd that M1 exo was internalized by SKOV3 cells.The 50 μg/ml M1 exo groups showed a significant reduction in SKOV3 cell migration,invasion and apoptosis,as well as decreased p-JAK1 and p-STAT3 protein expression,compared to the PBS group(t=4.08～21.63,P<0.05).The 100 μg/ml M1 exo group further suppressed SKOV3 cell proliferation,migration,invasion and apoptosis,and lowers p-JAK1 and p-STAT3 expression,compared to the 50 μg/ml group(t=3.61～15.60,P<0.05).Following co-culture,the supernatant of both M1 exo groups demonstrated a marked increase in IL-6 and TNF-α levels and the immune killing rate of NK-92MI cells,compared to the PBS group(t=21.76～14.88,P<0.05).Notably,the 100 μg/ml M1 exo group exhibited the highest levels of IL-6,TNF-α and NK-92MI immune killing rate,with statistically significant differences(t=4.41～6.88,P<0.05).Conclusions M1 macrophage ex-tracellular vesicles may inhibit SKOV3 cell proliferation,migration,invasion,and immune escape by downregulating the JAK1/STAT3 signaling pathway,promoting cell apoptosis.
陈淑贤;钱红霞;吕晓芳
石家庄市妇幼保健院妇一科,石家庄 050000石家庄市妇幼保健院妇一科,石家庄 050000石家庄市妇幼保健院妇一科,石家庄 050000
医药卫生
卵巢癌M1型巨噬细胞外泌体免疫逃逸酪氨酸激酶1/信号转导和转录激活因子3通路
ovarian cancerM1 type macrophagesextracellular vesiclesimmune escapeJAK1/STAT3 pathway
《现代检验医学杂志》 2026 (1)
24-28,5
河北省卫健委2024年度医学科学研究课题(编号:20242378).
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