miR-200c/PAI-2 promotes the progression of triple negative breast cancer via M1/M2 polarization induction of macrophage

Ziqi Meng; Rui Zhang; Yixuan Wang; Guang Zhu; Tiefeng Jin; Chunguo Li; Songnan Zhang

doi:10.1016/j.intimp.2019.106028

miR-200c/PAI-2 promotes the progression of triple negative breast cancer via M1/M2 polarization induction of macrophage

Int Immunopharmacol. 2020 Apr:81:106028. doi: 10.1016/j.intimp.2019.106028. Epub 2019 Dec 2.

Authors

Ziqi Meng¹, Rui Zhang¹, Yixuan Wang¹, Guang Zhu¹, Tiefeng Jin², Chunguo Li³, Songnan Zhang⁴

Affiliations

¹ Department of Pathology and Cancer Research Center, Yanbian University Medical College, Yanji 133002, China.
² Department of Pathology and Cancer Research Center, Yanbian University Medical College, Yanji 133002, China. Electronic address: jintf@ybu.edu.cn.
³ Department of Oncology, Jilin City Central Hospital, Jilin 132011, China. Electronic address: xinfeifusu@163.com.
⁴ Department of Oncology, Yanbian University Hospital, Yanji 133002, China. Electronic address: zhangsn21@163.com.

PMID: 31801690
DOI: 10.1016/j.intimp.2019.106028

Abstract

Purpose: To investigate the effect of miR-200c/PAI-2 on macrophage polarization into M2-type TAMs in TNBC.

Methods and materials: PAI-2 expression in MDA-MB-231^con, MDA-MB-231^miR-200ab and MDA-MB-231^miR-200c breast cancer cells was evaluated by RT-PCR and immunofluorescence (IF), while the expression of the TAM marker F4/80 and the M2-type TAM marker CD206 in MDA-MB-231^con, MDA-MB-231^miR-200c and MDA-MB-231^{miR-200c/siPAI-2} mouse lung metastatic tumor tissues was examined with immunohistochemistry (IHC). The effects of RAW264.7 cells on MDA-MB-231^con, MDA-MB-231^miR-200c and MDA-MB-231^{miR-200c/siPAI-2} were examined by transwell co-culture. CD206 expression in RAW264.7 cells were confirmed by immunostaining. The level of PAI-2 and IL-10 in the co-culture supernatants were assessed using ELISA.

Results: 1. RT-PCR and IF analysis showed that PAI-2 was upregulated in MDA-MB-231^miR-200c cells. 2. IHC assays analysis showed that the numbers of F4/80 and CD206 positive cells were increased in MDA-MB-231^miR-200c tumor tissues, while in MDA-MB-231^{miR-200c/siPAI-2} tumor tissues were decreased. 3. Transwell co-culture assays analysis showed that MDA-MB-231^miR-200c cells significantly promoted the cell migration ability compared with the control group, while knockdown PAI-2 significantly inhibited the cell migration ability (P < 0.05). 4. Transwell co-culture and immunostaining assays analysis showed that overexpression miR-200c in MDA-MB-231 cell line increased the CD206 expression in RAW264.7 cells, while knockdown PAI-2 decreased. 5. ELISA assays analysis showed that miR-200c-mediated MDA-MB-231 cells significantly increased the secretion of PAI-2 and IL-10, while decreased the secretion of PAI-2 and IL-10 in MDA-MB-231 ^{miR-200c/siPAI-2} cells.

Conclusions: miR-200c promotes the malignant progressions of TNBC by PAI-2 upregulation and M2 phenotype macrophages polarization.

Keywords: Cell migration; MDA-MB-231; Macrophages; PAI-2; Transwell co-culture; miR-200c.

MeSH terms

Animals
Cell Line, Tumor
Coculture Techniques
Disease Progression
Female
Gene Expression Regulation, Neoplastic / immunology
Gene Knockdown Techniques
Humans
Lectins, C-Type
Lung Neoplasms / genetics
Lung Neoplasms / immunology*
Lung Neoplasms / secondary
Macrophage Activation / genetics
Macrophage Activation / immunology
Macrophages / immunology*
Macrophages / metabolism
Mannose Receptor
Mannose-Binding Lectins
Mice
MicroRNAs / metabolism*
Plasminogen Activator Inhibitor 2 / genetics*
RAW 264.7 Cells
Receptors, Cell Surface
Triple Negative Breast Neoplasms / genetics
Triple Negative Breast Neoplasms / immunology
Triple Negative Breast Neoplasms / pathology*
Up-Regulation / immunology
Xenograft Model Antitumor Assays

Substances

Lectins, C-Type
MIRN200 microRNA, human
Mannose Receptor
Mannose-Binding Lectins
MicroRNAs
Plasminogen Activator Inhibitor 2
Receptors, Cell Surface