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

代朦; 靳鑫; 雷优扬; 明佳

doi:10.16016/j.1000-5404.201801107

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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

基础医学 | 更新时间：2026-01-09

- 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
- Effect of miR-382 overexpressing tumor-associated macrophages on biological properties of triple-negative breast cancer 4T1 cells
- 第三军医大学学报 2018年40卷第15期页码：1375-1382
- 作者机构：
  
  400010 重庆, 重庆医科大学附属第二医院三腺外科
- 作者简介：
  
  [ 明佳, E-mail:mjia1001@sina.com MING Jia, E-mail:mjia1001@sina.com
- 基金信息：
  
  重庆市渝中区科技计划项目(20160104);重庆市卫生计生委医学科研计划项目(2016ZDXM012)
- DOI：10.16016/j.1000-5404.201801107
  中图分类号： R394.3;R730.23;R737.9
- 收稿：2018-01-15，
  
  修回：2018-03-15，
  
  纸质出版：2018-08-15
- 稿件说明：
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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.

Meng DAI, Xin JIN, Youyang LEI, et al. Effect of miR-382 overexpressing tumor-associated macrophages on biological properties of triple-negative breast cancer 4T1 cells[J]. Journal of Third Military Medical University, 2018, 40(15): 1375-1382.
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.

Meng DAI, Xin JIN, Youyang LEI, et al. Effect of miR-382 overexpressing tumor-associated macrophages on biological properties of triple-negative breast cancer 4T1 cells[J]. Journal of Third Military Medical University, 2018, 40(15): 1375-1382. DOI： 10.16016/j.1000-5404.201801107.

摘要

目的

探讨microRNA-382（miR-382）在肿瘤相关巨噬细胞（tumor-associated macrophages，TAMs）中的表达对三阴性乳腺癌（triple-negative breast cancer，TNBC）细胞（4T1）生物学特性的影响。

方法

以小鼠腹腔巨噬细胞（mouse peritoneal macrophages，PMs）作为研究对象，分为PMs组，TAMs组和L-TAMs组；Western blot与qRT-PCR检测极化指标（IL-10、TNF-α）、miR-382、PGC-1α和NF-κB的表达变化；流式细胞术检测TAMs的极化变化（CD86、CD206）；激光共聚焦显微镜成像检测PGC-1α和NF-κB的表达变化；以4T1细胞作为研究对象，分为4T1组，4T1+PMs组和4T1+L-PMs组，流式细胞术检测细胞凋亡率与细胞周期；Transwell侵袭实验检测细胞侵袭能力。

结果

与PMs组比较，TAMs组IL-10、CD206和PGC-1α表达明显增加（

0.05），miR-382、TNF-α和CD86表达明显降低（

0.05），NF-κB转录活性显著降低（

0.05），而L-TAMs组TNF-α、CD86和miR-382表达明显增高（

0.05），IL-10、CD206和PGC-1α表达明显降低（

0.05），NF-κB转录活性显著增强（

0.05）；与4T1组比较，4T1+PMs组侵袭能力明显增强（

0.05），凋亡率明显下降（

0.05），而4T1+L-PMs组侵袭能力明显下降（

0.05），凋亡率明显增高（

0.05）。

结论

TAMs通过调节miR-382的表达可抑制TNBC的侵袭和增殖，其机制可能与TAMs中miR-382抑制PGC-1α的表达、增强NF-κB的转录活性，进而抑制TAMs的M2分化有关。

Abstract

Objective

To investigate the effect of microRNA-382 (miR-382) in tumor-associated macrophages (TAMs) on the biological characteristics of triple-negative breast cancer cells

in vitro

Methods

Mouse peritoneal macrophages (PMs) were used as research subjects and divided into PMs group

TAMs group and L-TAMs group (miR-382 overexpressing PMs cocultured with 10%TAM in a Transwell chamber system). Western blotting and qRT-PCR were used to detect the markers of polarization (IL-10

TNF-α) and the expression changes of miR-382

PGC-1α and NF-κB. Flow cytometry was employed to determine the polarization changes of TAMs (CD86

CD206)

and laser confocal microscopy to observe the expression of PGC-1α and NF-κB. Triple-negative breast cancer cells 4T1 were divided into 4T1 group

4T1+PMs co-culture group and 4T1+L-PMs co-culture group. Flow cytometry was adopted to detect the apoptotic rate and cell cycle. Transwell invasion assay was used to measure the invasion ability of the cells.

Results

Compared with the PMs group

the expression of IL-10

CD206 and PGC-1α was increased (

0.05)

and that of TNF-α

CD86 and miR-382 was decreased (

0.05)

transcriptional activity of NF-κB was reduced (

0.05) in the TAMs group; While in the L-TAMs

the results were totally opposite to those in the PMs group (

0.05). Compared with the 4T1 group

the invasive ability of 4T1+PMs group was increased (

0.05) and the apoptotic rate was decreased (

0.05)

while the invasive ability was lowered (

0.05) and the apoptotic rate was enhanced (

0.05) in the 4T1+L-PMs group.

Conclusion

TAMs inhibit the invasion and proliferation of triple-negative breast cancer by regulating the expression of miR-382

which may be related to the inhibition of the expression of PGC-1α by miR-382

the transcriptional activity of NF-κB and the inhibition of M2 differentiation of TAMs.

关键词

Keywords

references

COSTA R, SHAH A N, SANTA-MARIA C A, et al . Targeting epidermal growth factor receptor in triple negative breast cancer:new discoveries and practical insights for drug development[J ] . Cancer Treat Rev, 2017, 53:111-119. DOI:10.1016/j.ctrv.2016.12.010.

DENKERT C, LIEDTKE C, TUTT A, et al . Molecular alterations in triple-negative breast cancer-the road to new treatment strategies[J ] . Lancet, 2017, 389(10087):2430-2442. DOI:10.1016/S0140-6736(16)32454-0.

余腾骅, 王智亮, 赵晨晖, 等.雌激素激活GPR30/ERK通路促进三阴性乳腺癌MDA-MB-468细胞迁移及侵袭[J].第三军医大学学报, 2014, 36(20):2077-2082.

YU T H, WANG Z L, ZHAO C H, et al .Estrogen enhances migration and invasion in triple-negative breast cancer MDA-MB-468 cells via activating GPR30/ERK signaling pathway[J ] .J Third Mil Med Univ, 2014, 36(20):2077-2082..

YU T, DI G. Role of tumor microenvironment in triple-negative breast cancer and its prognostic significance[J]. Chin J Cancer Res, 2017, 29(3):237-252. DOI:10.21147/j.issn.1000-9604.2017.03.10.

HAO N B, LU M H, FAN Y H, et al . Macrophages in tumor microenvironments and the progression of tumors[J ] . Clin Dev Immunol, 2012, 2012:948098. DOI:10.1155/2012/948098.

FUJIMOTO H, SANGAI T, ISHⅡ G, et al . Stromal MCP-1 in mammary tumors induces tumor-associated macrophage infiltration and contributes to tumor progression[J ] . Int J Cancer, 2009, 125(6):1276-1284. DOI:10.1002/ijc.24378.

MAR-AGUILAR F, MENDOZA-RAMIREZ J A, MALAGON-SANTIAGO I, et al . Serum circulating microRNA profiling for identification of potential breast cancer biomarkers[J ] . Dis Markers, 2013, 34(3):163-169. DOI:10.32 33/DMA-120957.

CALIN G A, CROCE C M. MicroRNA signatures in human cancers[J]. Nat Rev Cancer, 2006, 6(11):857-866. DOI:10.1038/nrc1997.

DENARDO D G, BRENNAN D J, REXHEPAJ E, et al . Leukocyte complexity predicts breast cancer survival and functionally regulates response to chemotherapy[J ] . Cancer Discov, 2011, 1(1):54-67. DOI:10.1158/2159-8274.CD-10-0028.

YUAN Z Y, LUO R Z, PENG R J, et al . High infiltration of tumor-associated macrophages in triple-negative breast cancer is associated with a higher risk of distant metastasis[J ] . Onco Targets Ther, 2014, 7:1475-1480. DOI:10.2147/OTT.S61838.

MING J, ZHOU Y, DU J, et al . miR-381 suppresses C/EBP-dependent Cx43 expression in breast cancer cells[J ] . Biosci Rep, 2015, 35(6):e00266-e00266. DOI:10.1042/bsr20150167.

PINEDA-TORRA I, GAGE M, DE JUAN A, et al . Isolation, culture, and polarization of murine bone marrow-derived and peritoneal macrophages[J ] . Methods Mol Biol, 2015, 1339:101-109. DOI:10.1007/978-1-4939-2929-0_6.

COLEGIO O R, CHU N Q, SZABO A L, et al . Functional polarization of tumour-associated macrophages by tumour-derived lactic acid[J ] . Nature, 2014, 513(7519):559-563. DOI:10.1038/nature13490.

YAN W, LIU X, MA H, et al . Tim-3 fosters HCC development by enhancing TGF-β-mediated alternative activation of macrophages[J ] . Gut, 2015, 64(10):1593-1604. DOI:10.1136/gutjnl-2014-307671.

ZHANG B, WANG J, GAO J, et al . Alternatively activated RAW264.7 macrophages enhance tumor lymphangiogenesis in mouse lung adenocarcinoma[J ] . J Cell Biochem, 2009, 107(1):134-143. DOI:10.1002/jcb.22110.

COFFELT S B, TAL A O, SCHOLZ A, et al . Angiopoietin-2 regulates gene expression in TIE2-expressing monocytes and augments their inherent proangiogenic functions[J ] . Cancer Res, 2010, 70(13):5270-5280. DOI:10.1158/0008-5472.CAN-10-0012.

PONOMAREV E D, VEREMEYKO T, BARTENEVA N, et al . MicroRNA-124 promotes microglia quiescence and suppresses EAE by deactivating macrophages via the C/EBP-α-PU.1 pathway[J ] . Nat Med, 2011, 17(1):64-70. DOI:10.1038/nm.2266..

MENG Q L, LIU F, YANG X Y, et al . Identification of latent tuberculosis infection-related microRNAs in human U937 macrophages expressing Mycobacterium tuberculosis Hsp16.3[J ] . BMC Microbiol, 2014, 14:37. DOI:10.1186/1471-2180-14-37.

GRAFF J W, DICKSON A M, CLAY G, et al . Identifying functional microRNAs in macrophages with polarized phenotypes[J ] . J Biol Chem, 2012, 287(26):21816-21825. DOI:10.1074/jbc.M111.327031.

ZHANG Y, ZHANG M, ZHONG M, et al . Expression profiles of miRNAs in polarized macrophages[J ] . Int J Mol Med, 2013, 31(4):797-802.DOI:10.3892/ijmm.2013.1260.

PUIGSERVER P, WU Z, PARK C W, et al . A cold-inducible coactivator of nuclear receptors linked to adaptive thermogenesis[J ] . Cell, 1998, 92(6):829-839. DOI:10.1016/s0092-8674(00)81410-5.

LEBLEU V S, O'CONNELL J T, GONZALEZ HERRERA K N, et al . PGC-1α mediates mitochondrial biogenesis and oxidative phosphorylation in cancer cells to promote metastasis[J ] . Nat Cell Biol, 2014, 16(10):992-1003. DOI:10.1038/ncb3039.

MCGUIRK S, GRAVEL S P, DEBLOIS G, et al . PGC-1α supports glutamine metabolism in breast cancer[J ] . Cancer Metab, 2013, 1(1):22. DOI:10.1186/2049-3002-1-22.

ZHANG S, LIU X, LIU J, et al . PGC-1 alpha interacts with microRNA-217 to functionally regulate breast cancer cell proliferation[J ] . Biomed Pharmacother, 2017, 85:541-548. DOI:10.1016/j.biopha.2016.11.062.

CHEW C L, CONOS S A, UNAL B, et al . Noncoding RNAs:Master Regulators of Inflammatory Signaling[J ] . Trends Mol Med, 2018, 24(1):66-84. DOI:10.1016/j.molmed.2017.11.003.

FURRER R, EISELE P S, SCHMIDT A, et al . Paracrine cross-talk between skeletal muscle and macrophages in exercise by PGC-1α-controlled BNP[J ] . Sci Rep, 2017, 7:40789. DOI:10.1038/srep40789.

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