Co-culture of ovarian cancer stem-like cells with macrophages induced SKOV3 cells stemness via IL-8/STAT3 signaling
Co-culture of ovarian cancer stem-like cells with macrophages induced SKOV3 cells stemness via IL-8/STAT3 signaling
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DOI：
10.1016/j.biopha.2018.04.022
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发表时间：
2018-07-01
影响因子：
7.5
通讯作者：
Luo, Xin
中科院分区：
文献类型：
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作者：
Ning, Yingxia;Cui, Yinghong;Luo, Xin
在癌症生物学领域的最新概念中，肿瘤微环境与癌症干细胞高度相关，并且在肿瘤进展中起关键作用。本研究旨在探讨THP-1细胞来源的巨噬细胞与SKOV 3来源的卵巢癌干细胞样细胞（OCSLCs）共培养诱导SKOV 3细胞向干细胞分化的机制。评估球体形成、软琼脂集落形成以及CD 133和CD 44的表达水平，以反映OCSLC特性。ELISA用于评估与或不与SKOV 3衍生的OCSLC共培养的巨噬细胞中的分泌谱变化。对于机制评估，使用rhIL-8、IL-8中和抗体（IL-8 Ab）、信号转导子和转录激活子3（STAT 3）shRNA和STAT 3 cDNA。结果表明，与THP-1细胞共培养的巨噬细胞IL-10、VEGF、MMP-9、IL-8的分泌及CD 163和STAT 3的表达水平均较THP-1细胞增加，而IL-12和NO的表达水平则较THP-1细胞减少，反映了M2巨噬细胞的极化。向THP-1细胞条件培养基中加入rhIL-8促进SKOV 3细胞中M2巨噬细胞的极化和干性，而向共培养条件培养基中加入IL- 8 Ab则抑制了这一作用。STAT 3的过表达可诱导SKOV 3细胞M2期巨噬细胞的极化和干细胞化，而THP-1细胞中STAT 3的敲低可抑制这一效应。重要的是，STAT 3过表达挽救了IL-8 Ab对SKOV 3细胞中M2巨噬细胞极化和干性的影响。这些结果表明，与SKOV 3衍生的OCSLC共培养的巨噬细胞在SKOV 3细胞中诱导的干性涉及IL-8/STAT 3信号传导。
Among recent concepts in the cancer biology field, the tumor microenvironment is highly associated with cancer stem cells, and plays a key role in tumor progression. This study aimed to explore the mechanism that the stemness induction of SKOV3 cell line by macrophages derived from THP-1 cells, which was co-cultured with SKOV3-derived ovarian cancer stem-like cells (OCSLCs). Sphere formation, soft agar colony formation, and expression levels of CD133 and CD44 were assessed to reflect OCSLC properties. ELISA was used to evaluate secretion profile changes in macrophages co-cultured with or without SKOV3-derived OCSLCs. For mechanistic evaluation, rhIL-8, IL-8 neutralizing antibody (IL-8 Ab), signal transducer and activator of transcription 3 (STAT3) shRNA and STAT3 cDNA were used. The results showed that IL-10, VEGF, MMP-9, IL-8 secretion and CD163 and STAT3 expression levels in macrophages co-cultured with OCSLCs were increased compared with those from THP-1 cells, while IL-12 and NO amounts were significantly reduced, reflecting M2 macrophage polarization. Addition of rhIL-8 to THP-1 cell conditioned media promoted M2 macrophage polarization and stemness in SKOV3 cells, which were suppressed by IL- 8 Ab addition to co-culture conditioned media. Consistently, overexpression of STAT3 induced M2 macrophage polarization and stemness in SKOV3 cells, which were inhibited by STAT3 knockdown in macrophages from THP-1 cells. Importantly, STAT3 overexpression rescued the effects of IL-8 Ab on M2 macrophage polarization and stemness in SKOV3 cells. These results suggested that stemness induction in SKOV3 cells by macrophages co-cultured with SKOV3-derived OCSLCs involved IL-8/STAT3 signaling.