Department of Obstetrics and Gynecology, Cheil General Hospital and Women's Healthcare Center, Dankook university College of Medicine, Seoul, Republic of Korea.
Department of Obstetrics and Gynecology Guro Hospital, College of Medicine, Korea University, Seoul, Republic of Korea.
Int J Oncol. 2015 Oct;47(4):1451-9. doi: 10.3892/ijo.2015.3122. Epub 2015 Aug 13.
Epithelial-to-mesenchymal transition (EMT) facilitates the invasion and metastasis of cancer cells. EMT seems to be mediated by the interaction between cancer cells and human mesenchymal stem cells (hMSCs) in the tumor microenvironment. The present study is intended to identify specific cytokines as potent inducers of EMT associated hMSCs-tumor interactions. We used ovarian cancer cell lines (SKOV-3 and IGROV-1), endometrial cancer cell line (Ishikawa) and hMSCs (bone marrow MSC, amniotic membrane MSC and decidua MSC). The expressions of EMT markers (E-cadherin, Snail, Twist and N-cadherin) were analyzed using quantitative RT-PCR, immunofluorescence and western blot analysis. Matrix metalloproteinases (MMP-2 and MMP-9), Matrigel invasion assay, and wound healing assay were used to analyze cell migration and invasion. Gynecologic cancer cells directly co-cultured with hMSCs had contact-dependent altered morphology and growth patterns. IL-6 was elevated in all co-cultures using a human cytokine array. After IL-6 treatment of cancer cell lines, RT-PCR and western blot analysis indicated a decrease in an epithelial marker and an increase in mesenchymal markers. Also, cancer cells with IL-6 significantly increase in MMP-2 and MMP-9 and significantly enhance the migration ability compared to untreated cells (P<0.05), as shown by wound healing assay. On Matrigel invasion assay, treated cells displayed significantly increased invasiveness compared to untreated cancer cells. Gyneocologic cancer cells exposed to IL-6 acquired mesenchymal properties that facilitated metastasis and invasion by promoting EMT. The present study suggests that IL-6 of the tumor microenvironment has a critical role in oncogenic EMT.
上皮间质转化（EMT）促进癌细胞的侵袭和转移。EMT 似乎是由肿瘤微环境中癌细胞与人类间充质干细胞（hMSC）之间的相互作用介导的。本研究旨在鉴定特定细胞因子作为 EMT 相关 hMSC-肿瘤相互作用的有效诱导剂。我们使用卵巢癌细胞系（SKOV-3 和 IGROV-1）、子宫内膜癌细胞系（Ishikawa）和 hMSC（骨髓 MSC、羊膜 MSC 和蜕膜 MSC）。使用定量 RT-PCR、免疫荧光和 Western blot 分析分析 EMT 标志物（E-钙粘蛋白、Snail、Twist 和 N-钙粘蛋白）的表达。使用基质金属蛋白酶（MMP-2 和 MMP-9）、Matrigel 侵袭测定和划痕愈合测定分析细胞迁移和侵袭。妇科癌细胞与 hMSC 直接共培养具有接触依赖性的形态和生长模式改变。使用人细胞因子阵列检测到所有共培养物中的 IL-6 升高。在 IL-6 处理癌细胞系后，RT-PCR 和 Western blot 分析表明上皮标志物减少，间充质标志物增加。此外，与未经处理的细胞相比，用 IL-6 处理的癌细胞中 MMP-2 和 MMP-9 显著增加，并且迁移能力显著增强（P<0.05），如划痕愈合测定所示。在 Matrigel 侵袭测定中，处理过的细胞显示出比未经处理的癌细胞显著增加的侵袭性。暴露于 IL-6 的妇科癌细胞获得了促进 EMT 的间充质特性，从而促进了转移和侵袭。本研究表明，肿瘤微环境中的 IL-6 在致癌 EMT 中具有关键作用。
