ERK通路在胰腺癌细胞株SW1990吉西他滨化疗耐药中的作用  

The Role of the ERK Signaling Pathway in Gemcitabine Chemoresistance in Pancreatic Cancer Cell Line SW1990

作　　者：谢德荣[1] 郭双双[2] 杨琼[1] 江志敏[1] 陈邓林[1] 毕卓菲[1] 马雯[1] 

机构地区：[1]中山大学附属第二医院肿瘤科,广州市510120 [2]河南科技大学第一附属医院肿瘤科

出　　处：《中国肿瘤临床》2008年第14期829-832,共4页Chinese Journal of Clinical Oncology

基　　金：广东省科技计划项目基金资助(编号:2007B031516014)~~

摘　　要：目的:探讨细胞外信号调节激酶(ERK1/2)通路、多药耐药基因(mdr-1)、核糖核苷还原酶M1(RRM1),在胰腺癌细胞株SW1990吉西他滨(GEM)化疗耐药中的相互关系及作用。方法:通过浓度梯度递增法,诱导建立耐各种浓度GEM的胰腺癌细胞株。应用免疫组化图像分析方法,半定量检测各耐药细胞株中ERK1/2蛋白的表达;RT-PCR的方法检测mdr-1、RRM1mRNA表达,条带图使用Band凝胶分析软件进行半定量分析;MTT法测定OD492光密度值,计算IC50值。结果:建立的耐药细胞株可以在GEM终浓度分别为0、30、60、100、150、200nmol/L的培养液中稳定生长并传代。ERK1/2、mdr-1、RRM1的表达均随着耐GEM浓度的增加而升高,细胞耐GEM的浓度分别与mdr-1、RRM1基因表达呈现出高度正相关(r=0.960,P=0.002和r=0.966,P=0.002);ERK1/2灰度值也与mdr-1、RRM1基因的表达存在相关性(r=-0.943,P=0.005和r=-0.883,P=0.02)。耐GEM200nmol/L的细胞株,ERK1/2灰度值、mdr-1/β-actin、RRM1/β-actin分别为164.22±13.17、1.41±0.04、1.45±0.18;经阻断剂U0126作用后三者的表达均呈现同步降低,分别为186.85±13.14、0.23±0.02、0.21±0.03;而激活剂EGF作用后,其表达呈现升高趋势,分别为106.55±16.45、1.50±0.07、1.52±0.12。耐GEM0nmol/L和200nmol/L浓度的细胞株,其IC50值分别为4.104和10.20,经阻断剂U0126处理后,IC50值下降到3.26和4.50;而经EGF处理后,IC50值则分别上升到8.89和17.17。结论:ERK通路可能参与调控mdr-1、RRM1基因表达,从而介导胰腺癌细胞株SW1990吉西他滨化疗抵抗。Objective： To investigate the role of extracellular signal-regulated kinase pathway （ERK）, multidrug resistance 1 （MDR-1） and ribonucleoside reductase M1 （RRM1） in Gemcitabine （GEM） chemoresistance in the pancreatic cancer cell line SW1990. Methods： The GEM-resistant pancreatic cancer cell line model was constructed using a stepwise increase in concentration gradient of Gemcitabine. Immunocytochemistry was performed to detect the expression of ERK1/2 in the established cell lines in a semiquantitative way, and the mRNA expression of MDR-1 and RRM1 in the GEM-resistant pancreatic cancer cell lines was detected by RT-PCR. MTT assay was used to determine the IC50. Results： The established pancreatic cancer cell lines cultured in medium with GEM at different concentrations （0, 30, 60, 100, 150 and 200 nmol/L） were able to grow and maintain their resistance through several passages. The expression of ERK1/2, MDR-1 and RRM1 were elevated according to the increasing GEM concentration. There was a highly positive correlation between MDR-1 or RRM1 expression and GEM concentration （r=0.960,P=0.002 and r=0.966, P=-0.002）. The gray scale of ERKI/2 was also correlated with the expression of MDR-1 and RRM1 （r=-0.943, P=0.005 and r=-0.883, P=0.02）. At a concentration of 200 nmol/L, the gray scale of ERK1/2, MDR-1/β-actin, and RRM1/β-actin of the GEM-resistant pancreatic cancer cell line was 164.22±13.17, 1.41 ±0.04 and 1.45±0.18, respectively. The expression of these genes decreased synchronously after the cells were treated with ERK1/2 signaling pathway specific inhibitor U0126 to the gray scale values of 186.85±13.14, 0.23±0.02 and 0.21±0.03, respectively. Conversely, there was a tendency to decreased expression of these genes after the cells were treated with epidermal growth factor （EGF）. The gray scale values ascended to 106.55±16.45, 1.5±0.07 and 1.52±0.12, respectively. The IC50 of the GEM-resistant pancreatic cancer cell lines at the concentration of 0 nmol/L and 200 nmol

关 键 词：细胞外信号调节激酶 胰腺肿瘤 吉西他滨 耐药性 

分 类 号：R735.9[医药卫生—肿瘤] R743.3[医药卫生—临床医学]