ERK/MAP激酶抑制剂U0126增强索拉菲尼对原代胃癌细胞的抑制效应

U0126 (ERK/MAP kinase inhibitor) Enhances the Suppression of Sorafenib Against Primary Gastric Cancer Cells

目的:研究ERK/MAP激酶抑制剂U0126是否增强索拉菲尼对原代胃癌细胞的抑制效应,并探索其可能机制。方法:选取病理确诊胃癌患者外科手术标本培养原代胃癌细胞。使用索拉菲尼和U0126分别或同时处理原代细胞。MTT试验检测处理后细胞增殖情况;流式细胞术检测细胞凋亡情况;Q-PCR和Western Blotting分别检测转录水平和翻译水平增殖和凋亡相关分子的表达情况。结果:使用索拉菲尼和U0126同时处理的原代胃癌细胞,其增殖明显受到抑制,凋亡增加,并均强于单独处理组。同时处理组中p-ERK和p-Raf表达水平降低,Bim、Bax和Bad水平升高,水平变化均强于单独处理组,P<0.01。结论:U0126能够增强索拉菲尼抑制胃癌细胞增殖和促进细胞凋亡的效应,其机制可能是二者在抑制ERK/MAPK通路及诱导Bcl-2家族中促凋亡分子表达上有协同效应。

Objective： To investigate whether ERK/MAP kinase inhibitor U0126 enhances the suppression of Sorafenib against primary gastric cancer cells and explore the possible mechanism（s）.Methods： The surgically resected specimens pathologically diagnosed to be gastric cancer were selected for primary cell culture.And the primary cells were treated with Sorafenib or/and U0126.MTT assay was applied to detect the proliferation of cells;flow cytometry for detection of cell apoptosis;Q-PCR and Western Blotting to detect expression of related molecules.Results： Cells treated with Sorafenib and U0126 showed significant suppression of cell proliferation and promotion of cell apoptosis compared with that treated with Sorafenib or U0126 alone.Additionally,the treatment with these two reagents rapidly inhibited expression of p-ERK and p-Raf,and increase of expression of Bim,Bax,and Bad was observed,as well（P0.01）.Conclusion： U0126 can enhance the effect of Sorafenib on cell proliferation and apoptosis,the mechanism of which may be associated with the synergistic effect displayed by Sorafenib and U0126 on suppression of ERK/MAPK signal passway and induction of anti-apoptotic proteins of Bcl-2 family.