M1型巨噬细胞对小鼠膀胱癌MB49细胞活力、迁移、侵袭及凋亡的影响  

Effects of M1 macrophages on the viability, migration, invasion and apoptosis of mouse bladder cancer MB49 cells

作　　者：张暑军 李青青 乔春林 勾璇 王新敏[2] 章乐[1] ZHANG Shu-jun;LI Qing-qing;QIAO Chun-lin;GOU Xuan;WANG Xin-min;ZHANG Le(Department of Pathophysiology/Xinjiang Key Laboratory of Endemic and Ethnic Diseases,School of Medicine,Shihezi University,Shihezi 832000,China;Department of Urology,First Affiliated Hospital,School of Medicine,Shihezi Universi-ty,Shihezi 832008,China)

机构地区：[1]石河子大学医学院病理生理学教研室/新疆地方病与民族高发病教育部重点实验室,新疆石河子832000 [2]石河子大学医学院第一附属医院泌尿外科,新疆石河子832008

出　　处：《中国病理生理杂志》2022年第1期1-10,共10页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81660330);新疆地方与民族高发病教育部重点实验室开放基金(No.KF2021-5);石河子大学科研项目(No.ZZZC20261A);石河子大学医学院第一附属医院重点基金(No.ZD202006)。

摘　　要：目的:探究共培养体系下M1型巨噬细胞对小鼠膀胱癌MB49细胞活力、迁移、侵袭及凋亡的影响。方法:分别以0、50、100和200μg/L的脂多糖(LPS)刺激小鼠RAW264.7巨噬细胞24 h后,采用免疫荧光技术、Western blot及流式细胞术检测M1型巨噬细胞标志物CD86和诱导型一氧化氮合酶的表达情况。建立RAW264.7-MB49细胞共培养体系,分为MB49组、M0+MB49组和M1+MB49组,12 h后收集MB49细胞,CCK-8法检测各组MB49细胞活力,划痕实验和Transwell实验检测各组MB49细胞的迁移和侵袭能力,Hoechst 33258染色检测各组MB49细胞的凋亡情况,ELISA检测培养上清液中白细胞介素6(IL-6)浓度。结果:筛选出了诱导RAW264.7巨噬细胞向M1亚型极化的最佳LPS浓度和作用时间分别为100μg/L和24 h。与MB49组及M0+MB49组相比,M1+MB49组中MB49细胞的活力及迁移和侵袭能力均显著下降(P<0.05),凋亡细胞显著增多(P<0.05),培养上清液中IL-6浓度显著升高(P<0.05)。结论:在共培养体系中M1型巨噬细胞可抑制小鼠膀胱癌MB49细胞的活力、迁移和侵袭,并促进其凋亡。AIM:To explore the effect of M1 macrophages on the viability,migration,invasion and apoptosis of mouse bladder cancer MB49 cells in a co-culture system.METHODS:After stimulating mouse RAW264. 7 macrophages with 0,50,100 and 200 μg/L lipopolysaccharide(LPS)for 24 h,the expression of M1 macrophage markers(CD86 and inducible nitric oxide synthase)was detected by immunofluorescence,Western blot and flow cytometry. The RAW264. 7-MB49 cell co-culture system was established and divided into 3 groups:MB49 group(control group),M0+MB49 group and M1+MB49 group. MB49 cells were collected after 12 h of co-culture. CCK-8 assay was used to measure the viability of MB49 cells. The migration and invasion capacities of MB49 cells were detected by scratch test and Transwell test. The apoptosis of MB49 cells was observed by Hoechst 33258 staining. The concentration of interleukin-6(IL-6)in the culture supernatant was detected by ELISA.RESULTS:The best LPS concentration for inducing macrophages into M1 type was 100 μg/L in our study. Compared with control group and M0+MB49 group,the viability,and migration and invasion capacities of MB49 cells were significantly reduced,while the apoptosis of MB49 cells was significantly increased in M1+MB49 group(P<0. 05). The concentration of IL-6 in M1+MB49 group was significantly higher than that in the other two groups(P<0. 05).CONCLUSION:In the co-culture system,M1 macrophages could inhibit the viability,migration and invasion,but promote the apoptosis of mouse bladder cancer MB49 cells.

关 键 词：肿瘤微环境 M1型巨噬细胞 膀胱癌 细胞迁移 细胞侵袭 

分 类 号：R737.14[医药卫生—肿瘤] R730.23[医药卫生—临床医学]