摘要
目的:探究THP-1来源的肿瘤相关巨噬细胞(TAMs)对宫颈癌SiHa细胞迁移和侵袭的影响及其相关分子机制。方法:佛波脂(PMA)和IL-4将THP-1细胞诱导为TAMs,流式细胞仪检测其表面CD204和CD206分子的表达;将TAMs与SiHa细胞共培养,采用划痕实验、Transwell实验检测TAMs对SiHa细胞迁移和侵袭能力的影响;免疫印迹法检测共培养后SiHa细胞上皮间质转化相关蛋白的变化,RT-PCR检测转录因子Snail/Slug mRNA表达水平。结果:PMA作用于THP-1细胞24 h,IL-4继续作用72 h后,细胞表面CD204和CD206分子表达明显增加,TAMs诱导成功;与未共培养组相比,共培养组SiHa细胞迁移和侵袭能力显著增强(P<0.001);共培养组SiHa细胞E-cadherin蛋白表达水平显著降低,N-cadherin、Vimentin蛋白、Snail/Slug mRNA表达水平显著升高(P<0.05)。结论:TAMs可能通过上调转录因子Snail的表达促进SiHa细胞上皮间质转化进程,从而促进其侵袭和转移。
Objective:To explore the influence and related molecular mechanism of tumor-associated macrophages (TAMs)from THP-1 on the migration and invasion in cervical cancer SiHa cells.Methods:THP-1 cells were induced by phorbol myristate acetate (PMA)and interleukin-4 (IL-4)as TAMs.The expression of CD204 and CD206 on their surface were determined by flow cytometry.TAMs were co-cultured with SiHa cells.The effects of TAMs on the abilities of migration and invasion of SiHa cells were detected by Scratch test and Transwell assay.The changes of epithelial-mesenchymal transition related proteins in SiHa cells were detected by Western blot after co-culture.The mRNA expression levels of transcription factor Snail/Slug were detected by RT-PCR.Results:After 24 hours of being treated with PMA and 72 hours with IL-4, the expression of CD204 and CD206 on the surface of THP-1 cells were significantly increased.The induction of TAMs was proved to be successful.Compared with the non-culture group, both the migratory ability and the invasive ability of SiHa cells in the co-culture group were significantly enhanced (P<0.001).The protein expression level of E-cadherin remarkably decreased in the co-culture group.The protein expression levels of N-cadherin, Vimentin and the mRNA expression levels of Snail/Slug significantly increased(P<0.05).Conclusion:TAMs may promote the process of epithelial-mesenchymal transition of SiHa cells by up-regulating the expression of transcription factor Snail, thus promoting the invasion and metastasis of SiHa cells.
作者
吉紫阳
任云青
JI Zi-Yang;REN Yun-Qing(Fenyang College of Shanxi Medical University,Fenyang 032200, China)
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2020年第4期451-456,共6页
Chinese Journal of Immunology
基金
山西医科大学汾阳学院科研项目基金(No.1422,No.2019C08)资助
关键词
肿瘤相关巨噬细胞
宫颈癌
迁移
侵袭
上皮间质转化
Tumor-associated macrophages
Cervical cancer
Migration
Invasion
Epithelial-mesenchymal transition
