miR-382-5p affects polarization of tumor-associated macrophages in breast cancer through Akt/mTOR signaling pathway

Hua ZHOU; Yuanyuan WANG; Zijing LIN; Jia MING

doi:10.16016/j.1000-5404.202102001

您当前的位置：

首页 >

文章列表页 >

miR-382-5p affects polarization of tumor-associated macrophages in breast cancer through Akt/mTOR signaling pathway

Basic Medicine | 更新时间：2026-01-09

- miR-382-5p affects polarization of tumor-associated macrophages in breast cancer through Akt/mTOR signaling pathway
- Journal of Third Military Medical University Vol. 43, Issue 14, Pages: 1358-1365(2021)
- 作者机构：
  
  400010 重庆，重庆医科大学附属第二医院乳甲外科
- 作者简介：
- 基金信息：
  
  the "Kuanren Elite" Backbone Talent Project of the Second Affiliated Hospital of Chongqing Medical University(KY2019G016);the General Project of Technology Innovation in Application Demonstration and the Social and Livelihood of Chongqing(CSTC2018jscx-msybx0020);the Project of Science and Technology Plan of Yuzhong District, Chongqing(20160104)
- DOI：10.16016/j.1000-5404.202102001
  CLC： R394.3;R730.23;R737.9
- Received：01 February 2021，
  
  Revised：2021-04-27，
  
  Published：30 July 2021
- 稿件说明：
移动端阅览
Hua ZHOU, Yuanyuan WANG, Zijing LIN, et al. miR-382-5p affects polarization of tumor-associated macrophages in breast cancer through Akt/mTOR signaling pathway[J]. Journal of Third Military Medical University, 2021, 43(14): 1358-1365.
DOI：

Hua ZHOU, Yuanyuan WANG, Zijing LIN, et al. miR-382-5p affects polarization of tumor-associated macrophages in breast cancer through Akt/mTOR signaling pathway[J]. Journal of Third Military Medical University, 2021, 43(14): 1358-1365. DOI： 10.16016/j.1000-5404.202102001.

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(

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(

0.05)；Akt/mTOR信号通路相关蛋白磷酸化水平明显降低(

0.05)；4T1细胞的增殖、侵袭、迁移能力被抑制(

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

Objective

To 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.

Methods

Breast 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.

Results

The level of miR-382-5p in breast cancer TAMs was significantly lower than that in macrophages of pairing adjacent tissues (

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 (

0.05)

and the phosphorylation levels of the proteins related to Akt/mTOR signaling pathway were reduced (

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 (

0.05).

Conclusion

miR-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.

关键词

Keywords

references

KOMOHARA Y, FUJIWARA Y, OHNISHI K, et al . Tumor-associated macrophages: potential therapeutic targets for anti-cancer therapy[J ] . Adv Drug Deliv Rev, 2016, 99(Pt B): 180-185. DOI:10.1016/j.addr.2015.11.009.

MANTOVANI A, MARCHESI F, MALESCI A, et al . Tumour-associated macrophages as treatment targets in oncology[J ] . Nat Rev Clin Oncol, 2017, 14(7): 399-416. DOI:10.1038/nrclinonc.2016.217.

PINTON L, MASETTO E, VETTORE M, et al . The immune suppressive microenvironment of human gliomas depends on the accumulation of bone marrow-derived macrophages in the center of the lesion[J ] . J Immunother Cancer, 2019, 7(1): 58. DOI:10.1186/s40425-019-0536-x.

WEI C, YANG C, WANG S, et al . Crosstalk between cancer cells and tumor associated macrophages is required for mesenchymal circulating tumor cell-mediated colorectal cancer metastasis[J ] . Mol Cancer, 2019, 18(1): 64. DOI:10.1186/s12943-019-0976-4.

TANG X, MO C, WANG Y, et al . Anti-tumour strategies aiming to target tumour-associated macrophages[J ] . Immunology, 2013, 138(2): 93-104. DOI:10.1111/imm.12023.

HONKANEN T J, TIKKANEN A, KARIHTALA P, et al . Prognostic and predictive role of tumour-associated macrophages in HER2positive breast cancer[J ] . Sci Rep, 2019, 9(1): 10961. DOI:10.1038/s41598-019-47375-2.

EDELMAN M J, WANG X, HODGSON L, et al . Phase Ⅲ randomized, placebo-controlled, double-blind trial of celecoxib in addition to standard chemotherapy for advanced non-small-cell lung cancer with cyclooxygenase-2 overexpression: CALGB 30801 (Alliance)[J ] . J Clin Oncol, 2017, 35(19): 2184-2192. DOI:10.1200/JCO.2016.71.3743.

BUTOWSKI N, COLMAN H, DE GROOT J F, et al . Orally administered colony stimulating factor 1 receptor inhibitor PLX3397 in recurrent glioblastoma: an ivy foundation early phase clinical trials consortium phase Ⅱ study[J ] . Neuro Oncol, 2016, 18(4): 557-564. DOI:10.1093/neuonc/nov245.

REID T, ORONSKY B, SCICINSKI J, et al . Safety and activity of RRx-001 in patients with advanced cancer: a first-in-human, open-label, dose-escalation phase 1 study[J ] . Lancet Oncol, 2015, 16(9): 1133-1142. DOI:10.1016/S1470-2045(15)00089-3.

NASSAR F J, NASR R, TALHOUK R. MicroRNAs as biomarkers for early breast cancer diagnosis, prognosis and therapy prediction[J]. Pharmacol Ther, 2017, 172: 34-49. DOI:10.1016/j.pharmthera.2016.11.012.

LIU G, ABRAHAM E. MicroRNAs in immune response and macrophage polarization[J]. Arterioscler Thromb Vasc Biol, 2013, 33(2): 170-177. DOI:10.1161/ATVBAHA.112.300068.

ESSANDOH K, LI Y, HUO J, et al . MiRNA-mediated macrophage polarization and its potential role in the regulation of inflammatory response[J ] . Shock, 2016, 46(2): 122-131. DOI:10.1097/SHK.0000000000000604.

FENG J, QI B, GUO L, et al . miR-382 functions as a tumor suppressor against esophageal squamous cell carcinoma[J ] . World J Gastroenterol, 2017, 23(23): 4243-4251. DOI:10.3748/wjg.v23.i23.4243.

WANG J, CHEN C, YAN X, et al . The role of miR-382-5p in glioma cell proliferation, migration and invasion[J ] . OncoTargets Ther, 2019, 12: 4993-5002. DOI:10.2147/OTT.S196322.

ZHENG S, LI M, MIAO K, et al . SNHG1 contributes to proliferation and invasion by regulating miR-382 in breast cancer[J ] . Cancer Manag Res, 2019, 11: 5589-5598. DOI:10.2147/CMAR.S198624.

杨芳, 伍菲凡, 王晓燕, 等. Ficoll和Percoll法分离结直肠癌相关巨噬细胞的比较[J]. 实用医学杂志, 2015(9): 1394-1398. DOI:10.3969/j.issn.1006-5725.2015.09.004.

YANG F, WU F F, WANG X Y, et al . A comparative study of tumor-associated macrophages isolation in colorectal carcinoma by Ficoll-Hy-paque density gradient centrifugation and Percoll density gradient centrifugation[J ] . J Prac Med, 2015(9): 1394-1398. DOI:10.3969/j.issn.1006-5725.2015.09.004.

代朦, 靳鑫, 雷优扬, 等. 过表达miR-382的肿瘤相关巨噬细胞对三阴性乳腺癌生物学特性的影响[J]. 第三军医大学学报, 2018, 40(15): 1375-1382. DOI:10.16016/j.1000-5404.201801107.

DAI M, JIN X, LEI Y Y, et al . Effect of miR-382 overexpressing tumor-associated macrophages on biological properties of triple-negative breast cancer 4T1 cells[J ] . J Third Mil Med Univ, 2018, 40(15): 1375-1382. DOI:10.16016/j.1000-5404.201801107.

靳鑫, 倪田根, 王宁, 等. PGC-1α通过线粒体介导巨噬细胞极化状态的机制研究[J]. 第三军医大学学报, 2019, 41(1): 56-62. DOI:10.16016/j.1000-5404.201808139.

JIN X, NI T G, WANG N, et al . Mechanism of mitochondria-mediated PGC-1α in macrophage polarization[J ] . J Third Mil Med Univ, 2019, 41(1): 56-62. DOI:10.16016/j.1000-5404.201808139.

BANERJEE S, XIE N, CUI H, et al . MicroRNA let-7c regulates macrophage polarization[J ] . J Immunol, 2013, 190(12): 6542-6549. DOI:10.4049/jimmunol.1202496.

WANG Z, BRANDT S, MEDEIROS A, et al . MicroRNA 21 is a homeostatic regulator of macrophage polarization and prevents prostaglandin E2-mediated M2 generation[J ] . PLoS ONE, 2015, 10(2): e0115855. DOI:10.1371/journal.pone.0115855.

LI Y, ZHAO L, SHI B, et al . Functions of miR-146a and miR-222 in tumor-associated macrophages in breast cancer[J ] . Sci Rep, 2015, 5: 18648. DOI:10.1038/srep18648.

POPOVA N V, JUCKER M. The role of mTOR signaling as a therapeutic target in cancer[J]. Int J Mol Sci, 2021, 22(4): 1743. DOI:10.3390/ijms22041743.

VERGADI E, IERONYMAKI E, LYRONI K, et al . Akt signaling pathway in macrophage activation and M1/M2 polarization[J ] . J Immunol, 2017, 198(3): 1006-1014. DOI:10.4049/jimmunol.1601515.

MA Q, HOU L, GAO X, et al . NKAP promotes renal cell carcinoma growth via AKT/mTOR signalling pathway[J ] . Cell Biochem Funct, 2020, 38(5): 574-581. DOI:10.1002/cbf.3508..

PAJEROWSKI A G, NGUYEN C, AGHAJANIAN H, et al . NKAP is a transcriptional repressor of notch signaling and is required for T cell development[J ] . Immunity, 2009, 30(5): 696-707. DOI:10.1016/j.immuni.2009.02.011.

CHEN D, LI Z, YANG Q, et al . Identification of a nuclear protein that promotes NF-kappaB activation[J ] . Biochem Biophys Res Commun, 2003, 310(3): 720-724. DOI:10.1016/j.bbrc.2003.09.074.

ZHOU D, HUANG C, LIN Z, et al . Macrophage polarization and function with emphasis on the evolving roles of coordinated regulation of cellular signaling pathways[J ] . Cell Signal, 2014, 26(2): 192-197. DOI:10.1016/j.cellsig.2013.11.004.

Views

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

Mechanism of mitochondria-mediated PGC-1α in macrophage polarization

A novel estrogen receptor expression stratification (low/moderate/high) based on Chinese population provides more accurate prognosis prediction for HR⁺/HER2^- early breast cancer patients

Sulfasalazine antagonizes boosted metastasis of breast cancer induced by ionizing radiation

Effect of transmembrane protein 196 on migration and invasion of breast cancer

Effect of circular RNA hsa_circ_0000231 interacting with HnRNPK on proliferation, migration, and apoptosis in breast cancer

Related Author

Xin JIN

Tiangen NI

Ning WANG

Haojun LUO

Youyang LEI

WANG Chengfang

ZHOU Yuqin

ZHANG Yanling

Related Institution

Department of Breast and Thyroid Surgery， First Affiliated Hospital， Army Medical University （Third Military Medical University）

State Key Laboratory of Trauma, Burns and Combined Injury, Department of Anti-Radiation Medicine, Chongqing Engineering Research Center for Nanomedicine, Institute of Combined Injury, Faculty of Military Preventive Medicine, Army Medical University (Third Military Medical University)

Department of Environmental Health, College of Military Preventive Medicine, Army Medical University (Third Military Medical University)

Institute of Toxicology, College of Military Preventive Medicine, Army Medical University (Third Military Medical University)

Department of Breast and Thyroid Surgery, Army Medical Center of PLA

⁰