Genistein与5-FU通过p42/44 MAPK通路调控Bax诱导肝癌细胞凋亡  

Genistein and 5-FU inhibit apoptosis of human hepatocellular cancer cells by regulating Bax via p42/44 MAPK pathway

作　　者：赵忠新 迟大鹏[2] 王钰粟 梅庆步[3] 于海涛[3] 岳丽玲[3] 王玉[3] 王秀华[3] 刘丹[3] 

机构地区：[1]齐齐哈尔市建华医院普外科,黑龙江省齐齐哈尔市161006 [2]上海中医药大学附属岳阳医院神经外科,上海市200437 [3]齐齐哈尔医学院基础医学院,黑龙江省齐齐哈尔市161006

出　　处：《世界华人消化杂志》2014年第14期2003-2007,共5页World Chinese Journal of Digestology

基　　金：黑龙江省教育厅科学技术研究基金资助项目;No.12531792~~

摘　　要：目的:探讨在染料木黄酮(genistein,Gen)与氟尿嘧啶(5-fluorouracil,5-FU)诱导肝癌MHCC97-L细胞凋亡作用中p42/44丝裂原活化蛋白激酶(mitogen-activated protein kinases,MAPKs)对Bax的调控.方法:倒置显微镜观察细胞密度和细胞形态;荧光显微镜(Annexin V-FITC/PI)检测细胞凋亡及细胞死亡;Western blot检测Bax蛋白表达.结果:倒置显微镜下,所有药物组细胞密度均下降,死亡细胞及碎片散落在培养液中,贴壁细胞皱缩或变形,胞浆透明度下降,可见空泡结构,其中以PD98059+Gen+5-FU组变化最明显;荧光显微镜下,所有药物组均可检测到凋亡和死亡细胞;Western blot检测各组Bax蛋白的相对表达量由高到低依次为Gen+5-F U、G e n、5-F U、P D98059+G e n+5-F U、PD98059+5-FU、PD98059+Gen、PD98059、对照组,PD98059+Gen组的Bax蛋白表达与P D98059组十分接近,且明显低于G e n组;PD98059+5-FU组和PD98059+Gen+5-FU组的Bax蛋白表达介于单用抑制剂组与未用抑制剂预处理的对应药物组之间.结论:Gen、5-FU和Gen+5-FU均能诱导MHCC97-L细胞凋亡,并与上调Bax有关;Gen对Bax的调控与p42/44 MAPK通路关系密切;5-FU和联用组调控Bax需要有p42/44 MAPK通路参与.AIM： To explore whether genistein and 5-FU induce apoptosis of human hepatic cancer MHCC97-L cells by regulating Bax via the p42/44 MAPK pathway.METHODS： MHCC97-L cells were treated with genistein, 5-FU, PD98059, genistein ＋ 5-FU, PD98059 ＋ genistein, PD98059 ＋ 5-FU, or PD98059 ＋ genistein ＋ 5-FU, respectively. The density and morphology of MHCC97-L cells were observed under an inverted microscope. Apoptosis and cell death were detected after annexin V-FITC/PI staining under a fluorescence microscope. The expression of Bax protein was detected by Western blot.RESULTS： All drug groups showed decreased cell density, cell death and debris, cell shrinkage or deformation, reduced cytoplasmic transparency, and vacuoles structure within the ceils, with PD98059 ＋ genistein ＋ 5-FU group having the most obvious changes. Apoptotic cells and dead cells were detected in all drug groups. The relative expression of Bax protein was the highest in the genistein ＋ 5-FU group, followed by the genistein, 5-FU, PD98059 ＋ genistein ＋ 5-FU, PD98059 ＋ 5-FU, PD98059 ＋ genistein, PD98059 and control groups. The expression of Bax protein in the PD98059 ＋ genistein group was very close to that in the PD98059 group, but was significantly lower than that in the genistein group. CONCLUSION： Genistein and 5-FU can induce the apoptosis of MHCC97-L cells and increase Bax expression possibly by regulating Bax mainly through the p42/44 MAPK pathway.

关 键 词：丝裂原活化蛋白激酶 染料木黄酮 氟尿嘧啶 细胞凋亡 BAX 肝癌 

分 类 号：R735.7[医药卫生—肿瘤]