miR-200c/PAI-2 promotes the progression of triple negative breast cancer via M1/M2 polarization induction of macrophage
miR-200c/PAI-2 promotes the progression of triple negative breast cancer via M1/M2 polarization induction of macrophage
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miR-200c/PAI-2通过诱导巨噬细胞M1/M2极化促进三阴性乳腺癌进展
DOI：
10.1016/j.intimp.2019.106028
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发表时间：
2020-04-01
影响因子：
5.6
通讯作者：
Zhang, Songnan
中科院分区：
文献类型：
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作者：
Meng, Ziqi;Zhang, Rui;Zhang, Songnan
目的：为了研究miR-200 c/派-2对TNBC中巨噬细胞极化成M2型TAM的影响。派-2在MDA-MB-231（con）、MDA-MB-231（miR-200ab）和MDA-MB-231通过RT-PCR和免疫荧光（IF）评估（miR-200 c）乳腺癌细胞，而TAM标志物F4/80和M2型TAM标志物CD 206在MDA-MB-231（con）、MDA-MB-231（miR-200 c）和MDA-MB-231（miR-200 c/siPAI-2）小鼠肺转移肿瘤组织中的表达用免疫组织化学（IHC）检测。通过transwell共培养检测RAW264.7细胞对MDA-MB-231（con）、MDA-MB-231（miR-200 c）和MDA-MB-231（miR-200 c/siPAI-2）的影响。RAW264.7细胞中的CD 206表达通过免疫染色证实。ELISA法检测细胞培养上清中派-2和IL-10的水平。RT-PCR和IF分析显示派-2在MDA-MB-231（miR-200 c）细胞中表达上调。2.免疫组化分析显示，MDA-MB-231（miR-200 c/siPAI-2）肿瘤组织中F4/80和CD 206阳性细胞数增加，而MDA-MB-231（miR-200 c/siPAI-2）肿瘤组织中F4/80和CD 206阳性细胞数减少。3. Transwell共培养分析显示，与对照组相比，MDA-MB-231（miR-200 c）细胞迁移能力明显增强，而派-2敲低组细胞迁移能力明显受到抑制（P <0. 05）。4. Transwell共培养和免疫组化分析显示，MDA-MB-231细胞中过表达miR-200 c可增加RAW 264. 7细胞中CD 206的表达，而敲低派-2则降低其表达。5.结论：miR-200 c通过上调派-2和M2表型巨噬细胞极化促进TNBC的恶性进展。
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.