肿瘤相关成纤维细胞通过p38/MAPK信号通路促进人乳腺癌细胞迁移

Cancer-associated fibroblast（CAF） promotes migration of human breast cancer cells via activating the p38/MAPK signaling pathway

摘要:

目的：探讨肿瘤相关成纤维细胞（cancer-associated fibroblast，CAF）对人乳腺癌细胞BT549和MDA-MB-231的迁移能力影响及可能的分子机制。方法：CAF上清液作用人乳腺癌细胞BT549和MDA-MB-231后，采用Transwell迁移实验检测BT549和MDA-MB-231细胞的迁移能力变化，蛋白质印迹法检测迁移相关蛋白E-cadherin、Vimentin的表达及p38/MAPK通路的活化情况。CAF上清液处理人乳腺癌细胞BT549和MDA-MB-231后，采用SB203580抑制p38/MAPK通路，Transwell迁移实验和蛋白质印迹分别检测BT549和MDA-MB-231细胞的迁移能力及E-cadherin、Vimentin蛋白的表达变化。结果：CAF作用后乳腺癌细胞BT549和MDA-MB-231的穿膜细胞数增多（P＜0.05），抑制BT549和MDA-MB-231细胞中E-cadherin表达水平（P＜0.01），上调Vimentin的表达水平（P＜0.05）；CAF激活乳腺癌细胞BT549和MDA-MB-231 p38/MAPK信号通路，促进通路关键分子p-P38蛋白表达（P＜0.01）；SB203580抑制p38/MAPK通路，CAF作用后乳腺癌细胞BT549和MDA-MB-231穿膜细胞数明显减少（P＜0.01），且上调E-cadherin蛋白表达水平（P＜0.05）、下调Vimentin蛋白表达水平（P＜0.01）。结论：CAF通过激活p38/MAPK信号通路促进人乳腺癌细胞BT549和MDA-MB-231的迁移。

Abstract:

Objective：To investigate the effects of cancer-associated fibroblast（CAF） on migration of human breast cancer BT549 and MDA-MB-231 cells and its possible molecular mechanism. Methods：Effects of CAF on migration of BT549 and MDA-MB-231 were measured by transwell migration test after treated with CAF supernatant. Effects of CAF on migration related proteins E-cadherin and Vimentin were investigated by Western blot after treated with CAF supernatant. After treating the human breast cancer cell line BT549 and MDA-MB-231 with CAF supernatant，SB203580 was used to inhibit the p38/MAPK pathway，then Transwell migration assay and Western blot were used to detect the changes of migration and the expressions of E-cadherin and Vimentin in BT549 and MDA-MB-231 cells respectively. Results：CAF increased the number of migration cells in BT549 and MDA-MB-231 breast cancer cells（P<0.05），inhibited the expression of E-cadherin（P<0.01） and up-regulated the expression of Vimentin in BT549 and MDA-MB-231 cells（P<0.05）. CAF activated the p38 /MAPK signaling pathway of BT549 and MDA-MB-231 breast cancer cells and promoted the expression of p-P38 protein（P<0.01）. The ability of CAF to increase the number of migration cells in BT549 and MDA-MB-231 cells was significantly inhibited（P<0.01），and the expression of E-cadherin protein was up-regulated（P<0.05），while the expression of Vimentin protein was down-regulated（P<0.01） after SB203580 inhibited the p38/MAPK pathway. Conclusion：CAF promotes migration of human breast cancer BT549 and MDA-MB-231 cells via activating the p38/MAPK signaling pathway.