[1]周华,王园园,林子晶,等.miR-382-5p通过Akt/mTOR信号通路影响乳腺癌肿瘤相关巨噬细胞极化[J].陆军军医大学学报(原第三军医大学学报),2021,43(14):1358-1365.
 ZHOU Hua,WANG Yuanyuan,LIN Zijing,et al.miR-382-5p affects polarization of tumor-associated macrophages in breast cancer through Akt/mTOR signaling pathway[J].J Amry Med Univ (J Third Mil Med Univ),2021,43(14):1358-1365.
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miR-382-5p通过Akt/mTOR信号通路影响乳腺癌肿瘤相关巨噬细胞极化(/HTML )
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陆军军医大学学报(原第三军医大学学报)[ISSN:1000-5404/CN:51-1095/R]

卷:
43卷
期数:
2021年第14期
页码:
1358-1365
栏目:
基础医学
出版日期:
2021-07-30

文章信息/Info

Title:
miR-382-5p affects polarization of tumor-associated macrophages in breast cancer through Akt/mTOR signaling pathway
作者:
周华王园园林子晶明佳
重庆医科大学附属第二医院乳甲外科
Author(s):
ZHOU Hua WANG Yuanyuan LIN Zijing MING Jia

epartment of Breast and Thyroid Surgery, the Second Affiliated Hospital of Chongqing Medical University, Chongqing, 400010, China

关键词:
乳腺癌肿瘤相关巨噬细胞microRNAAkt/mTOR信号通路
Keywords:
breast cancer tumor-associated macrophages microRNA Akt/mTOR signaling pathway
分类号:
R394.3; R730.23; R737.9
文献标志码:
A
摘要:

目的探讨乳腺癌肿瘤相关巨噬细胞(tumor-associated macrophages,TAMs)中miR-382-5p的表达对巨噬细胞极化表型以及乳腺癌生物学特性的影响和可能机制。方法收集2018-2020年本院25例临床乳腺癌患者的肿瘤相关巨噬细胞及配对癌旁组织巨噬细胞,用qRT-PCR检测miR-382-5p的表达;利用miR-382-5p过表达慢病毒转染小鼠腹腔巨噬细胞(mouse peritoneal macrophages,PMs),qRT-PCR、Western blot检测巨噬细胞的M1/M2极化指标(iNOS、TNF-α、Arg-1、IL-10等)、miR-382-5p以及Akt/mTOR信号通路变化,采用流式细胞仪检测CD86、CD206的表达;将4T1细胞与TAMs、过表达miR-382-5p的TAMs共培养或培养上清液处理后,CCK-8实验检测4T1细胞的增殖能力,Transwell实验检测4T1细胞的侵袭能力,细胞划痕实验检测4T1细胞的迁移能力。结果25例临床标本的乳腺癌TAMs的miR-382-5p表达水平较配对癌旁组织巨噬细胞明显降低(P<0.05);与对照组比较,过表达miR-382-5p的TAMs的M1型极化标记物(iNOS、TNF-α、CD86等)的表达水平升高,M2型极化标记物(Arg-1、IL-10、CD206)的表达水平降低(P<0.05);Akt/mTOR信号通路相关蛋白磷酸化水平明显降低(P<0.05);4T1细胞的增殖、侵袭、迁移能力被抑制(P<0.05)。 结论miR-382-5p可能通过改变乳腺癌TAMs的极化状态影响乳腺癌细胞的侵袭、迁移等过程;其机制可能与miR-382-5p介导Akt/mTOR信号通路调节巨噬细胞极化相关。

Abstract:

ObjectiveTo investigate the effects of microRNA-382-5p (miR-382-5p) expression in breast cancer tumor-associated macrophages (TAMs) on the polarization phenotype of macrophages as well as the biological characteristics of breast cancer, and to explore its possible mechanism. MethodsBreast cancer TAMs and their paring macrophages from adjacent tissues of 25 breast cancer patients treated in our hospital between 2018 and 2020 were collected, and the expression of miR-382-5p was detected by qRT-PCR. Mouse peritoneal macrophages (PMs) were transfected with miR-382-5p over-expressed lentivirus. The changes of M1/M2 polarization indicators (iNOS, TNF-α, Arg-1, IL-10, etc.) and the expression of miR-382-5p and Akt/mTOR signaling pathway were determined by qRT-PCR and Western blotting, and the expression of CD86 and CD206 were tested using flow cytometry. After the 4T1 mouse breast cancer cells were co-cultured with TAMs and TAMs overexpressing miR-382-5p, respectively, or treated with the corresponding supernatants, the proliferative capacity of 4T1 cells was subsequently tested by CCK-8 assay, the invasion ability and the migration ability of 4T1 cells were observed using Transwell assay and cell scratch assay, respectively. ResultsThe level of miR-382-5p in breast cancer TAMs was significantly lower than that in macrophages of pairing adjacent tissues (P<0.05). Compared with the control group, the levels of M1-phenotype polarization markers (iNOS, TNF-α, CD86, etc.) were increased, while those of M2 indicators (Arg-1, IL-10, CD206) were decreased in the miR-382-5p overexpressed group (P<0.05), and the phosphorylation levels of the proteins related to Akt/mTOR signaling pathway were reduced (P<0.05). In addition, the abilities of proliferation, invasion and migration of 4T1 cells were all inhibited in the miR-382-5p overexpressed group as compared with the TAMs group (P<0.05). ConclusionmiR-382-5p may possibly play a vital role in the invasion and migration processes of breast cancer cells by affecting the polarization phenotypes of TAMs, and the mechanism may be associated with miR-382-5p mediated Akt/mTOR signaling pathway in the regulation of macrophage polarization.

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更新日期/Last Update: 2021-07-18