摘要
目的探究卵泡抑素相关基因3(follistatin-related gene 3,FSTL3)对肿瘤相关巨噬细胞(tumour-associated macro-phages,TAMs)在乳腺癌增殖、迁移和侵袭中的作用及其相关机制。方法收集2018年3月至2021年3月在海军军医大学第一附属医院21例乳腺癌患者的肿瘤及其癌旁组织,并提取巨噬细胞。使用佛波酯(phorbol-12-myristate-13-acetate,PMA)、PMA/干扰素-γ(interferon-γ,IFN-γ)和PMA/白细胞介素-4(interleukin-4,IL-4)诱导THP-1为初始、M1型和M2型巨噬细胞,分别为PMA组、PMA/IFN-γ组、PMA/IL-4组;采用脂质体转染法将si-FSTL3、si-NC转染至THP-1,再使用PMA/IL-4诱导转染后的THP-1,分别为si-FSTL3组、si-NC组。采用流式细胞术鉴定巨噬细胞分型。通过实时荧光定量聚合酶链式反应(quanlita-tive real time polymerase chain reaction,qRT-PCR)和Western blotting检测各组细胞FSTL3的mRNA和蛋白表达情况。收集各组巨噬细胞上清与购买的乳腺癌细胞系MCF7共孵育,通过细胞计数试剂盒(cell counting kit8,CCK8)检测MCF7细胞增殖能力,使用细胞划痕、Transwell实验检测MCF7细胞迁移和侵袭能力。结果与癌旁组织比较,乳腺癌患者TAMs的FSTL3基因表达增加,差异具有统计学意义(P<0.001),表型呈M2型极化。与PMA组相比,PMA/IL-4组高表达FSTL3,其上清共孵育的MCF7细胞增殖、迁移和侵袭能力增加,差异均具有统计学意义(P<0.001)。与si-NC组相比,si-FSTL3组向M2型极化能力减弱,其上清共孵育的MCF7细胞的增殖、迁移和侵袭能力减弱,差异具有统计学意义(P<0.05)。结论FSTL3通过正向调控TAMs向M2型极化,促进乳腺癌细胞增殖、迁移和侵袭。
Objective To explore the effects of FSTL3 on the tumor-associated macrophages(TAMs)in the proliferation,mi-gration and invasion of breast cancer and its related mechanism.Methods The tissue and paracancerous tissue of 21 patients with breast cancer who sought medical care in the First Affiliated Hospital of the Navy Medical University from March,2018 to March,2021 were recruited for study.Macrophages were extracted from tumors and adjacent tissues.Phorbol-12-myristate-13-acetate(PMA),interferon-γ(IFN-γ)and interleukin-4(IL-4)were used to induce initial M1 and M2 macrophages,which were designated as the PMA group,the PMA/IFN-γgroup and the PMA/IL-4 group.Si-FSTL3 and si-NC were transfected into THP-1 by liposome transfection method.Then,PMA/IL-4 was used to induce transfected THP-1,and assigned respectively as the si-FSTL3 group and the si-NC group.The macrophage typing was identified by flow cytometry.The expressions of Follistatin like 3(FSTL3)were detected by quantitative real time polymerase chain reaction(qRT-PCR)and Western blot.Macrophage supernatants of each group were collected and co-incubated with MCF7 cells.The proliferation,migration and invasion abilities of MCF7 cells were detected by cell counting kit8(CCK8),cell scratches and Transwell assay respectively.Results The expression of FSTL3 gene increased in TAMs,as com-pared with that of the paracancerous tissue group,with statistical significance(P<0.001).The phenotype tended to be M2 polariza-tion.Compared with the PMA group,the PMA/IL-4 group had higher expression of FSTL3,and the proliferation,migration and inva-sion abilities of MCF7 cells co-incubated with its supernatant increased,all with statistical significance(P<0.001).Compared with that of the si-NC group,the polarization ability to M2 type in the si-FSTL3 group weakened,and the proliferation,migration and inva-sion abilities of MCF7 cells co-incubated with the supernatant of si-FSTL3 group also weakened,also with statistical significance(P<0.005).Conclusion FSTL3 induces the proliferat
作者
吕凌云
彭宇
鲁亚琴
房贺
陈艳
姜玉
汪志平
Lyu Lingyun;Peng Yu;Lu Yaqin;Fang He;Chen Yan;Jiang Yu;Wang Zhiping(Health Management Center,Affiliated Hospital of Naval Medical University,Shanghai 200433,China)
出处
《海军医学杂志》
2022年第7期697-703,共7页
Journal of Navy Medicine
