Graduate Institute of Integrated Medicine and Chinese Medicine Research Center, China Medical University, No. 91, Hsueh-Shih Road, Taichung, 40402, Taiwan.
Graduate Institute of Biomedical Sciences, China Medical University, Taichung, Taiwan.
J Biomed Sci. 2022 Dec 22;29(1):109. doi: 10.1186/s12929-022-00888-x.
Ovarian cancer has the highest mortality among gynecological cancers due to late diagnosis and lack of effective targeted therapy. Although the study of interplay between cancer cells with their microenvironment is emerging, how ovarian cancer triggers signaling that coordinates with immune cells to promote metastasis is still elusive.
Microarray and bioinformatics analysis of low and highly invasive ovarian cancer cell lines were used to reveal periostin (POSTN), a matrix protein with multifunctions in cancer, with elevated expression in the highly invasive cells. Anchorage independent assay, Western blot, RNA interference, confocal analysis and neutralizing antibody treatment were performed to analyze the effects of POSTN on tumor promotion and to explore the underlying mechanism. Chemotaxis, flow cytometry and cytokine array analyses were undertaken to analyze the involvement of POSTN in cancer-associated fibroblast (CAF) and macrophage modulation. Correlations between POSTN expression levels and clinical characteristics were analyzed using the Oncomine, commercial ovarian cancer cDNA and China Medical University Hospital patient cohort. In vivo effect of POSTN on metastasis was studied using a mouse xenograft model.
Expression of POSTN was found to be elevated in highly invasive ovarian cancer cells. We observed that POSTN was co-localized with integrin β3 and integrin β5, which was important for POSTN-mediated activation of ERK and NF-κB. Ectopic expression of POSTN enhanced whereas knockdown of POSTN decreased cancer cell migration and invasion in vitro, as well as tumor growth and metastasis in vivo. POSTN enhanced integrin/ERK/NF-κB signaling through an autocrine effect on cancer cells to produce macrophage attracting and mobilizing cytokines including MIP-1β, MCP-1, TNFα and RANTES resulting in increased chemotaxis of THP-1 monocytes and their polarization to M2 macrophages in vitro. In agreement, tumors derived from POSTN-overexpressing SKOV3 harbored more tumor-associated macrophages than the control tumors. POSTN induced TGF-β2 expression from ovarian cancer cells to promote activation of adipose-derived stromal cells to become CAF-like cells expressing alpha smooth muscle actin and fibroblast activation protein alpha. Consistently, increased CAFs were observed in POSTN overexpressing SKOV3 cells-derived metastatic tumors. In clinical relevance, we found that expression of POSTN was positively correlated with advanced-stage diseases and poor overall survival of patients.
Our study revealed a POSTN-integrin-NF-κB-mediated signaling and its involvement in enhancing M2 macrophages and CAFs, which could potentially participate in promoting tumor growth. Our results suggest that POSTN could be a useful prognosis marker and potential therapeutic target.
卵巢癌是妇科癌症中死亡率最高的一种，因为其诊断较晚，缺乏有效的靶向治疗。尽管癌症细胞与其微环境之间相互作用的研究正在兴起，但卵巢癌如何引发与免疫细胞协调促进转移的信号仍然难以捉摸。
使用低侵袭性和高侵袭性卵巢癌细胞系的微阵列和生物信息学分析，揭示了具有多种癌症功能的基质蛋白——骨膜蛋白（POSTN）在高侵袭性细胞中表达升高。进行锚定非依赖性测定、Western blot、RNA 干扰、共焦分析和中和抗体处理，以分析 POSTN 对肿瘤促进的影响，并探讨其潜在机制。进行趋化性、流式细胞术和细胞因子阵列分析，以分析 POSTN 在癌症相关成纤维细胞（CAF）和巨噬细胞调节中的作用。使用 Oncomine、商业卵巢癌 cDNA 和中国医科大学附属医院患者队列分析 POSTN 表达水平与临床特征的相关性。使用小鼠异种移植模型研究 POSTN 对转移的体内影响。
发现 POSTN 在高侵袭性卵巢癌细胞中表达升高。我们观察到 POSTN 与整合素β3 和整合素β5 共定位，这对 POSTN 介导的 ERK 和 NF-κB 激活很重要。POSTN 的异位表达增强了，而 POSTN 的敲低减少了体外癌细胞的迁移和侵袭，以及体内肿瘤的生长和转移。POSTN 通过自分泌作用于癌细胞增强整合素/ERK/NF-κB 信号，产生吸引和动员巨噬细胞的细胞因子，包括 MIP-1β、MCP-1、TNFα 和 RANTES，导致 THP-1 单核细胞趋化性增加，并在体外向 M2 巨噬细胞极化。同样，来自 POSTN 过表达 SKOV3 的肿瘤比对照肿瘤含有更多的肿瘤相关巨噬细胞。POSTN 诱导卵巢癌细胞表达 TGF-β2，以促进脂肪来源的基质细胞激活成为表达α平滑肌肌动蛋白和成纤维细胞激活蛋白α的 CAF 样细胞。一致地，在 POSTN 过表达 SKOV3 细胞衍生的转移性肿瘤中观察到增加的 CAFs。在临床相关性方面，我们发现 POSTN 的表达与晚期疾病和患者总生存期不良呈正相关。
我们的研究揭示了 POSTN-整合素-NF-κB 介导的信号及其在增强 M2 巨噬细胞和 CAF 中的作用，这可能有助于促进肿瘤生长。我们的结果表明 POSTN 可以作为一个有用的预后标志物和潜在的治疗靶点。
