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

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。

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.