DADS诱导人胃癌细胞分化作用中ERK/AP-1通路的改变

Role of ERK/AP-1 pathway in the differentiation induced by DADS on human gastric caner cell

目的 探讨ERK/AP 1通路在二烯丙基二硫 (DADS)诱导人胃癌细胞分化中的作用 ,阐明DADS诱导人胃癌细胞分化的分子机制。方法 采用免疫细胞化学、图像分析及WesternBlot等方法 ,观察DADS作用于体外培养的人胃癌MGC80 3细胞前后AP 1成员c fos与c jun表达的改变以及ERKMAPK激酶的变化。结果 免疫细胞化学及图像分析结果显示 ,对照组c fos、c jun表达呈强阳性 ,而处理组呈阴性或弱阳性 ,阳性率明显降低 ,处理组光密度值较对照组明显低 (P <0 0 5 )。WesternBlot结果显示 :从 2 0、2 5、30到 35mg·L-1DADS处理癌细胞 ,DADS呈浓度依赖性抑制人胃癌细胞中ERK的活化 (P <0 0 5 ) ;并且DADS +MEK抑制剂PD980 5 9能完全阻断ERK活化 (P <0 0 5 ) ;时间效应上 ,在DADS刺激后 15～ 30min抑制作用最强 ,2h左右恢复至接近正常 (P <0 0 5 )。结论 ERK/AP

AIM To investigate the role of ERK/AP-1 pathway in the differentiation induced by diallyl disulfide (DADS) on human gastric cancer MGC803 cell. METHODS Immunocytochemical staining and morphometric quantitative analysis detected the expression of c-fos and c-jun;Western Blot which measured the activation of ERK was analyzed to elucidate the possible mechanism of DADS-induced human gastric cancer cell differentiation. RESULTS Immunocytochemical staining and morphometric quantitative analysis indicated that expression of c-fos and c-jun reduced(P<0.05).At 35 mg·L -1, DADS induced inhibition of activation of ERK in 15～30 minute;Simultaneous change of activation of ERK induced by DADS from 25 to 35 mg·L -1 showed a concentration-dependent manner(P<0.05). DADS-induced inhibition of activation of ERK was enhanced by PD98059, a specific MEK inhibitor(P<0.05). CONCLUSION Inhibition of ERK/AP-1 is involved in DADS induced differentiation in gastric cancer cells.