腫瘤相關(guān)巨噬細(xì)胞是由單核細(xì)胞在腫瘤細(xì)胞誘導(dǎo)下分化形成的,，通常具有M2型巨噬細(xì)胞表型。腫瘤相關(guān)巨噬細(xì)胞浸潤(rùn)在三陰性乳腺癌組織中較為常見(jiàn),，并且其豐度分布與預(yù)后不良相關(guān),，具體機(jī)制不詳。

　　2016年9月6日,，國(guó)際癌癥研究中心官方期刊《國(guó)際腫瘤學(xué)雜志》在線發(fā)表江西省南昌市第三醫(yī)院和復(fù)旦大學(xué)附屬腫瘤醫(yī)院丁景弦,、郭春根、胡平華,、陳軍,、劉秋明、吳曉波,、曹亞麗、吳炅的研究報(bào)告,，探討了腫瘤相關(guān)巨噬細(xì)胞在三陰性乳腺癌維系及其與乳腺癌轉(zhuǎn)移的關(guān)系,，并在此基礎(chǔ)上進(jìn)一步研究乳腺癌細(xì)胞對(duì)單核細(xì)胞定向分化的調(diào)節(jié)及其干預(yù)措施。

　　首先,，該研究通過(guò)分層隨機(jī)化收集復(fù)旦大學(xué)附屬腫瘤醫(yī)院三陰性乳腺癌組織標(biāo)本,，采用免疫組化技術(shù)分析M2型腫瘤相關(guān)巨噬細(xì)胞標(biāo)記CD163在乳腺癌中的表達(dá)分布。通過(guò)向培養(yǎng)基中加入集落刺激因子1（CSF1）人為誘導(dǎo)單核細(xì)胞株U937分化為M2型巨噬細(xì)胞,，采用鏡下觀察形態(tài),、RT-PCR及定量RT-PCR鑒定其表型,。分別用不同活細(xì)胞染料（CMTMR，CMFDA）標(biāo)記經(jīng)誘導(dǎo)分化的U937細(xì)胞及乳腺癌細(xì)胞,，利用體外促融合技術(shù)（聚乙二醇,，PEG1450）輔助融合，通過(guò)流式細(xì)胞儀熒光激活細(xì)胞分選（FACS）融合形成的雜交細(xì)胞,。通過(guò)體內(nèi)外實(shí)驗(yàn)比較融合細(xì)胞與其來(lái)源乳腺癌細(xì)胞相關(guān)生物學(xué)行為的差異,。

　　其次，該研究通過(guò)乳腺癌細(xì)胞與U937細(xì)胞共培養(yǎng)技術(shù)比較不同雌激素受體（ER）狀態(tài)乳腺癌細(xì)胞誘導(dǎo)單核細(xì)胞分化能力的差異,。為此,，該研究利用慢病毒載體系統(tǒng)藥物篩選了分別帶GFP-PURO和RFP-NEO標(biāo)記的U937細(xì)胞及乳腺癌細(xì)胞穩(wěn)定表達(dá)株，模擬單核細(xì)胞向腫瘤組織浸潤(rùn)的體內(nèi)過(guò)程,，研究不同乳腺癌細(xì)胞誘導(dǎo)單核細(xì)胞趨化遷移及侵襲能力,。采用Bio-Plex多重人細(xì)胞因子試劑盒檢測(cè)了不同ER狀態(tài)的乳腺癌細(xì)胞株單核細(xì)胞分化相關(guān)細(xì)胞因子CSF1的分泌情況。

　　結(jié)果發(fā)現(xiàn)：CD163在不同類(lèi)型乳腺癌組織中的表達(dá)存在差異,，在ER陰性乳腺癌標(biāo)本中表達(dá)較ER陽(yáng)性高,，而且CD163的表達(dá)高低是預(yù)后不良的獨(dú)立因素。經(jīng)過(guò)乙酸鹽（PMA）及CSF1的誘導(dǎo),，單核細(xì)胞株U937分化成M2型巨噬細(xì)胞,，表現(xiàn)在形態(tài)及免疫表型上表達(dá)CD163、CD204,。在PEG的輔助下分化的U937細(xì)胞與乳腺癌細(xì)胞株MDA-MB-231,、MDA-MB-468、MCF-7存在一定的融合現(xiàn)象,，經(jīng)FACS分選出來(lái)的雜交細(xì)胞在免疫表型及生物特性上同其來(lái)源細(xì)胞均有一定差異,。體外實(shí)驗(yàn)表明雜交細(xì)胞具有較強(qiáng)的遷移侵襲，而增殖能力較其來(lái)源細(xì)胞弱,。體內(nèi)實(shí)驗(yàn)研究表明,，雜交細(xì)胞成瘤能力及轉(zhuǎn)移能力較強(qiáng)。在乳腺癌細(xì)胞與U937細(xì)胞共培養(yǎng)誘導(dǎo)后者定向分化的研究中發(fā)現(xiàn)MDA-MB-231細(xì)胞能在較短時(shí)間（一周）內(nèi)將U937細(xì)胞定向分化為具有M2表型的腫瘤相關(guān)巨噬細(xì)胞,，而MCF-7細(xì)胞不具有短期誘導(dǎo)U937細(xì)胞分化的能力,。通過(guò)定量PCR及Bio-Plex多重人細(xì)胞因子試劑盒檢測(cè)發(fā)現(xiàn)MDA-MB-231與MCF-7在誘導(dǎo)單核細(xì)胞分化的相關(guān)細(xì)胞因子CSF1表達(dá)上存在明顯差異，這一結(jié)果與其在誘導(dǎo)U937細(xì)胞分化的功能上完全吻合,。

Int J Oncol. 2016 Sep 6. [Epub ahead of print]

CSF1 is involved in breast cancer progression through inducing monocyte differentiation and homing.

Ding J, Guo C, Hu P, Chen J, Liu Q, Wu X, Cao Y, Wu J.

The Third Hospital of Nanchang, Nanchang, Jiangxi 330009, P.R. China; Fudan University Shanghai Cancer Center, Shanghai 200032, P.R. China.

Despite the great progress in breast cancer research and treatment, measures for efficient targeting of triple-negative breast cancer (TNBC) are still lacking. The well-established dependency of cancer cells on their microenvironment suggests that targeting the tumor niche might form a novel therapeutic approach. We identified the tumor-associated macrophage (TAM) infiltration in breast cancer tissues by immunohistochemistry, and analyzed overall survival (OS). U937 co-cultures with MDA-MB-231, MDA-MB-468 and MCF-7, respectively, to simulate in vivo cellular interactions were assessed. In hormone-independent breast cancer cell conditioned media (CM), U937 differentiates into M2 macrophage as identified by morphological changes and expression of specific surface antigens CD163 and CD204. Moreover, MDA-MB-231 recruits U937, and colony-stimulating factor 1 (CSF1) level in MDA-MB-231 and MDA-MB-468 CM is much higher than that of MCF-7. Overexpression of CSF1 in MCF-7 fails to rebuild its aggressiveness both in vitro and in vivo since CSF1 was not found extracellularly, while genetic inhibition of CSF1 in MDA-MB-231 abrogates TAM infiltration and consequently reduces tumorigenesis in non-obese diabetic/severe combined immunodeficient (NOD/SCID) mice. Using various strategies we demonstrate that CSF1-induced TAMs specifically support breast cancer progression. Importantly, our results may reveal the efficacy of using targeted therapy against tumor niche and indicate that CSF1 inhibition may limit some breast cancer progression.

PMID: 27599777

DOI: 10.3892/ijo.2016.3680