白介素-13对与乳腺癌细胞共培养的成纤维细胞 SDF-1和 EGF 表达影响

摘要

Objective:The effects of IL-13 ( Interleukin-13 ) on SDF-1 ( Stromal cell derived factor 1 ) and EGF ( Epidermal growth factor) expression in fibroblasts co-cultured with breast cancer cells were investigated to explore the mechanism for IL-13 in the development of breast cancer.Methods:The co-culture of human breast cancer cell line MDA-MB-231 with the human skin fibroblast line CCC-ESF-1 ( ESF) was used in vitro and in tumor-burdened nude mice.The effects of IL-13 on SDF-1 and EGF expression in the co-cultured fibroblasts in vitro were analyzed using reverse transcription quantitative polymerase chain reaction ( RT-qPCR ) , flow cytometry and Western blot assay.The proliferation of the co-cultured human breast cancer cells in vitro was detected by Cell counting kit-8(CCK-8).The effects of IL-13 on SDF-1 and EGF expression in the fibroblasts of tumor tissue of tumor-burdened nude mice were analyzed using immunofluorescence and laser confocal microscope, and the tumor volumes were examined.Results: IL-13 could up-regulate SDF-1 and EGF expression in the fibroblasts co-cultured with breast cancer cells in vitro,and promoted the proliferation of the co-cultured breast cancer cells.In tumor-burdened nude mice,IL-13 enhanced SDF-1 and EGF expression of fibroblasts in tumor tissue, and accelerated tumor growth.Conclusion:IL-13 up-regulates SDF-1 and EGF expression of fibroblasts co-cultured with breast cancer cells.The molecular mechanism of promoting effect of IL-13 on breast cancer relates to SDF-1 and EGF of fibroblasts in breast cancer stroma.%目的：为了探索白细胞介素-13（Interleukin-13，IL-13）在乳腺癌发展中的作用机制，我们研究了IL-13对与乳腺癌细胞共生长的成纤维细胞表达SDF-1（Stromal cell derived factor 1）和EGF（Epidermal growth factor）的影响。方法：采用体外和荷瘤裸鼠体内人乳腺癌细胞株MDA-MB-231和人成纤维细胞株CCC-ESF-1（ESF）共培养的方法，用定量PCR（RT-qPCR）方法、流式细胞术和Western blot 方法检测在IL-13作用下体外共培养的成纤维细胞SDF-1和EGF 的表达，细胞增殖实验Cellcounting kit-8（CCK-8）观察IL-13对体外共培养的乳腺癌细胞增殖的影响；免疫荧光激光共聚焦显微镜观察在IL-13作用下荷瘤裸鼠体内与乳腺癌细胞共生长的成纤维细胞SDF-1和EGF 的表达，检测IL-13对荷瘤裸鼠肿瘤体积的影响。结果：IL-13上调体外与乳腺癌细胞共培养的成纤维细胞SDF-1和EGF 的表达，并促进共培养的乳腺癌细胞的增殖；IL-13上调荷瘤裸鼠乳腺癌组织成纤维细胞SDF-1和EGF 的表达，并促进荷瘤裸鼠肿瘤生长。结论：IL-13上调与乳腺癌细胞共培养的成纤维细胞SDF-1和EGF 的表达，IL-13对乳腺癌促进作用的分子机制涉及乳腺癌基质成纤维细胞的SDF-1和EGF。