表皮生长因子介导的NF-κB促进胰腺癌细胞MMP-9表达及侵袭的实验研究  

Epidermal Growth Factor-Mediated NF-κB Activity Induces Matrix Metalloproteinase-9 Expression and Invasion in Pancreatic Cancer Cell

作　　者：刘宇[1] 张浩[2] 杨野[1] 王欣[1] 李铁民[1] 郭仁宣[2] 

机构地区：[1]中国医科大学附属第一医院干诊科,沈阳110001 [2]中国医科大学附属第一医院普外科,沈阳110001

出　　处：《中国普外基础与临床杂志》2009年第8期603-608,共6页Chinese Journal of Bases and Clinics In General Surgery

摘　　要：目的观察表皮生长因子(EGF)对胰腺癌细胞增殖、黏附及侵袭力的影响及其对核因子-κB(NF-κB)和细胞基质金属蛋白酶(MMPs)表达的影响,探讨EGF促进胰腺癌侵袭的相关机理。方法通过细胞增殖、黏附及侵袭实验,观察在EGF影响下胰腺癌细胞黏附、增殖及侵袭能力变化;通过RT-PCR、MMPs明胶酶谱分析、Western blot及EMSA,检测胰腺癌细胞的MMP-2和MMP-9 mRNA及其蛋白表达以及NF-κB活性变化;并观察NF-κB抑制物吡咯烷二硫代氨基甲酸盐(PDTC)对EGF诱导的胰腺癌细胞NF-κB活性、MMP-9 mRNA及其蛋白表达以及肿瘤细胞侵袭力的变化。结果EGF能够增强胰腺癌细胞的侵袭能力,具有剂量依赖性(P<0.05)。EGF对胰腺癌细胞的黏附力及增殖力无明显影响。EGF处理后,肿瘤细胞的MMP-9蛋白和mRNA表达明显升高,EGF浓度为50 ng/ml时,肿瘤细胞的MMP-9表达最强,但对MMP-2蛋白和mRNA的表达则无影响。随着EGF浓度的增加,NF-κB活性增强,具有浓度依赖性(P<0.05)。与EGF单独处理相比,经PDTC和EGF共同处理后的胰腺癌细胞NF-κB活性与MMP-9蛋白和mRNA表达均降低;PDTC和EGF共同处理后的胰腺癌细胞侵袭力较EGF单独处理和对照均减弱(P<0.05)。结论EGF通过激活NF-κB的活性,诱导MMP-9表达,促进胰腺癌细胞的侵袭,而这一过程可以被PDTC抑制。Objective To observe the effect of epidermal growth factor （EGF） on the proliferation, adhesion, invasiveness and the activation of nuclear factor-κB （NF-κB）, matrix metalloproteinases （MMPs） expression and explore related mechanisms in pancreatic cancer cells. Methods Cell invasion assay, proliferation assay and adhesion assay were used to examine the proliferation, adhesion and invasiveness of pancreatic cancer cells, respectively. NF-κB activity was detected by electrophoretic mobility shift assay （EMSA）, and MMPs protein and mRNA expressions were investigated by gelatin zymography, Western blot and reverse transcriptase-polymerase chain reaction （RT-PCR）. Results EGF increased the invasiveness of pancreatic cancer cell in a dose-dependent manner （P〈 0.05）, but did not affect cell proliferation or adhesion. The expressions of MMP-9 mRNA and protein significantly increased after induction by EGF and were highest when EGF concentration was 50 ng/ml, while there was no effect on the expressions of MMP-2 mRNA and protein. Furthermore, NF-κB activity increased with increased concentration of EGF in a concentration-dependent manner （P〈0.05）. In addition, NF-κB activity and the expressions of MMP-9 mRNA and protein by pretreatment with both pyrrolidine dithiocarbamate （PDTC） and EGF decreased when compared that by pretreatment with EGF alone. The invasiveness of pancreatic cancer cell by pretreatment with both PDTC and EGF decreased when compared that by pretreatment with EGF alone and nothing （P〈0.05）.Conclusion The findings indicate that the NF-κB-mediated MMP-9 induction is essential for EGF-induced invasiveness in pancreatic cancer cells, which can be inhibited by PDTC.

关 键 词：表皮生长因子 胰腺癌 细胞基质金属蛋白酶 核因子-ΚB 侵袭 转移 

分 类 号：R735.9[医药卫生—肿瘤]