Department of Immunology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand.
Department of Gynecology and Obstetrics, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand.
Int J Oncol. 2021 May;58(5). doi: 10.3892/ijo.2021.5194. Epub 2021 Mar 2.
The tumor microenvironment composed of a mixture of stromal cells and their secretions has a marked impact on cancer progression. In particular, soluble factors and metabolites contribute to malignancy through the dysregulation of autophagy in cancer cells. The present study investigated the effects of ovarian cancer‑associated fibroblasts (OVCAFs) with their secretory substances on the autophagy and migration of ovarian cancer cells. The conditioned‑medium (CM) of OVCAFs isolated from fresh human ovarian cancer tissues was analyzed for the levels of 27 common cytokines/chemokines using a cytokine array. Autophagy in cancer cells was assessed by determining the expression of the vacuolar form of LC3 by western blot analysis and immunofluorescence. Cancer cell migration was assessed by Transwell migration assay. Interleukin (IL)‑8 was found to be the most highly upregulated cytokine among the cytokines/chemokines found in the OVCAF‑CM. The role of IL‑8 in ovarian cancer cell migration and its mechanistic link with autophagy was investigated. Recombinant human IL‑8 (rhIL‑8) stimulated the migration of SKOV3 and Kuramochi ovarian cancer cells, and concurrently downregulated basal autophagy, in concentration‑dependent manner. Compared to the CM of control counterpart normal fibroblasts isolated from benign ovaries (OVNF‑CM), the CM from 3 OVCAF isolates (namely, OVCAF‑9, ‑20 and ‑43) exerted effects similar to rhIL‑8 on both cancer cell lines. The pharmacological induction of autophagy with rapamycin or metformin attenuated the pro‑migratory effects of IL‑8. Neutralizing anti‑IL‑8 antibody counteracted the inhibitory effect of OVCAF‑CM on basal autophagy. On the whole, the present study highlights the involvement of IL‑8 released by CAFs in the ovarian tumor microenvironment in promoting cancer cell migration through the suppression of autophagy.
肿瘤微环境由基质细胞及其分泌物的混合物组成，对癌症的进展有显著影响。特别是，可溶性因子和代谢物通过调节癌细胞中的自噬作用导致恶性转化。本研究探讨了卵巢癌相关成纤维细胞（OVCAFs）及其分泌物质对卵巢癌细胞自噬和迁移的影响。使用细胞因子阵列分析从新鲜人卵巢癌组织中分离的 OVCAFs 的条件培养基（CM）中 27 种常见细胞因子/趋化因子的水平。通过 Western blot 分析和免疫荧光法测定空泡型 LC3 的表达来评估癌细胞中的自噬。通过 Transwell 迁移测定评估癌细胞迁移。细胞因子/趋化因子中发现白细胞介素（IL）-8 是上调最明显的细胞因子。研究了 IL-8 在卵巢癌细胞迁移中的作用及其与自噬的机制联系。重组人 IL-8（rhIL-8）以浓度依赖性方式刺激 SKOV3 和 Kuramochi 卵巢癌细胞的迁移，并同时下调基础自噬。与从良性卵巢（OVNF-CM）分离的对照正常成纤维细胞 CM 相比，来自 3 个 OVCAF 分离株（即 OVCAF-9、-20 和-43）的 CM 对这两种癌细胞系均具有类似于 rhIL-8 的作用。使用雷帕霉素或二甲双胍诱导自噬可减弱 IL-8 的促迁移作用。中和抗 IL-8 抗体抵消了 OVCAF-CM 对基础自噬的抑制作用。总的来说，本研究强调了 CAFs 释放的 IL-8 参与卵巢肿瘤微环境，通过抑制自噬来促进癌细胞迁移。
