Shiraz Institute for Cancer Research, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran.
Department of Immunology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran.
Cell Biol Int. 2018 Mar;42(3):334-343. doi: 10.1002/cbin.10905. Epub 2018 Jan 10.
Adipose-derived mesenchymal stem cells (ASCs) are known to have immunomodulatory properties through soluble factors or by direct cell-to-cell contact. This study aimed to assess the expression of HLA-G and IDO activity in breast cancer and normal ASCs and to see whether ASC is capable of modulating both tumor cells and immune system cells in vitro. ASCs were enzymatically isolated from 15 breast cancer patients and 10 normal individuals. Then they were cultured, and the impact of their conditioned media on the movement of the MDA-MB-231 breast cancer cell line was studied in wound healing scratch assay. Next, PBLs from the peripheral blood of normal individuals were separated and co-cultured with breast cancer and normal ASCs. PBLs proliferation and apoptosis were assessed using CFSE labeling dye and annexin V/7AAD staining, respectively. IDO activity and HLA-G protein expression in ASCs were examined using kynurenine assay and Western blotting, respectively. Tumor-derived ASCs, especially those from higher stages of breast cancer, have stronger effects on the proliferation and movement of MDA-MB-231 cells than normal ASCs (P-value < 0.05). Apoptosis in PBLs increased in the presence of ASCs compared to PBLs cultured alone (P-value < 0.05). In contrast, necrosis of PBLs decreased in the presence of ASCs compared to apoptosis in these cells (P-value < 0.001). Collectively, ASCs may have strategic effects on both tumor cells and cells of the immune system in the tumor microenvironment, resulting in tumor development, growth, and metastasis.
脂肪间充质干细胞 (ASCs) 通过可溶性因子或直接细胞间接触具有免疫调节特性。本研究旨在评估乳腺癌和正常 ASC 中 HLA-G 的表达和 IDO 活性，并观察 ASC 是否能够在体外调节肿瘤细胞和免疫系统细胞。通过酶从 15 名乳腺癌患者和 10 名正常人中分离 ASC，然后培养，并在划痕愈合试验中研究其条件培养基对 MDA-MB-231 乳腺癌细胞系运动的影响。接下来，从正常人的外周血中分离 PBL 并与乳腺癌和正常 ASC 共培养。使用 CFSE 标记染料和 annexin V/7AAD 染色分别评估 PBL 的增殖和凋亡。使用犬尿氨酸测定法和 Western blot 分别检测 ASC 中的 IDO 活性和 HLA-G 蛋白表达。与正常 ASC 相比，源自乳腺癌较高阶段的肿瘤衍生 ASC 对 MDA-MB-231 细胞的增殖和运动具有更强的作用（P 值 < 0.05）。与单独培养的 PBL 相比，PBL 中 ASC 的存在会增加细胞凋亡（P 值 < 0.05）。相比之下，与 PBL 凋亡相比，PBL 中 ASC 的存在会降低细胞坏死（P 值 < 0.001）。总之，ASC 可能在肿瘤微环境中对肿瘤细胞和免疫系统细胞都具有策略性影响，从而导致肿瘤的发展、生长和转移。
