缺氧对胰腺癌PANC1细胞增殖、凋亡及迁移的影响  

Effects of cobalt chloride mimetic hypoxia on the proliferation, apoptosis and migration of human pancreatic cancer cell line

作　　者：朱亮[1,2] 徐胜南[1] 庞慧芳[1] 赵慧贞[1] 覃华[1] 黎培员[1] 李德民[1] 赵秋[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院消化内科,武汉430030 [2]南昌大学第一附属医院消化内科

出　　处：《中华胰腺病杂志》2012年第2期111-114,共4页Chinese Journal of Pancreatology

摘　　要：目的探讨氯化钴（CoCl2）模拟缺氧对人胰腺癌细胞株PANC1增殖、凋亡和迁移的影响。方法分别用终浓度为0（对照）、100、200、400、800txmol／LCoCl：处理PANC1细胞24h，采用实时定量PCR、蛋白质印迹法分别检测细胞缺氧诱导因子（HIF）-1α、血管内皮生长因子（VEGF）mRNA及蛋白的表达，采用CCK-8法检测细胞增殖，流式细胞术检测细胞凋亡，细胞划痕试验检测细胞迁移能力。结果对照组及100、200、400、800μmol／LCoC12处理组HIF-1αmRNA表达量分别为1、1．08±0．12、1．12±0．09、1．04±0．11、0．66±0．07；VEGFmRNA表达量分别为1、2．69±0．35、4．81±0．54、2．19±0．21、0．79±0.08；HIF-1α蛋白表达量为0．23±0．03、0．36±0．04、1．15±0．11、1．08±0．09、0．44．±0.04；VEGF蛋白表达量为0．14±0．02、0．12±0．01、0．95±0．09、0．87．±0.09、0．55．±0.06；细胞存活率分别为100％、（98．43±2．88）％、（76．15±0．70）％、（53．87±0．77）％、（35．23±0．67）％；细胞凋亡率分别为（5．2±1．12）％、（5．74．±1．07）％、（6．82±1．85）％、（12．09±3．53）％、（31．88±6．95）％；细胞爬行距离分别为（43．24±3．67）％、（59．46±5．39）％、（80．56±8．05）％、（63．89±5．96）％、（9．09±1．59）％。与对照组相比较，处理组细胞VEGFmRNA、VEGF及HIF-1α蛋白表达、细胞爬行距离均呈先上升后下降的趋势（P〈0．05），HIF—l仪mRNA表达、细胞增殖率呈剂量依赖性降低，细胞凋亡率呈剂量依赖性升高。结论CoCl2达到一定浓度时可显著抑制PANC1细胞增殖，促进细胞凋亡；CoCl2对PANC1细胞迁移效应的双向作用与高浓度CoCl2对细胞直接损伤有关。Objective To investigate the influence of cobalt chloride （ CoCl2 ） -mimetic hypoxia on the proliferation, apoptosis and migration of human pancreatic cancer cell line PANC1. Methods PANC1 cells were treated with 0（control）, 100, 200, 400, 800 μmol/L CoCl2 respectively for 24 h. Real-time RT-PCR and Western blot were used to determine hypoxia induced factor （HIF）-lct mRNA and protein expression respectively, and cell counting kit-8 （CCK-8） assays, flow cytornetry and cell scratch test were used to examine the proliferation, apoptosis and migration of PANC1 cells, respectively. Results In the control group and 100, 200,400 and 800 Ixmol/L COC1-2 groups, the expressions of HIF-1α mRNA were 1, 1.08 ±0.12, 1.12 ± 0. 09, 1.04 ± 0. 11, 0.66 ± 0.07, and the expressions of VEGF mRNA were 1, 2.69 ± 0.35, 4.81 ± 0. 54, 2.19 ± 0. 21, 0.79 ± 0.08, while the expressions of HIF-1 α protein were 0.23 ± 0.03, 0. 36 ± 0. 04, 1.15 ± 0.11, 1.08 ±0.09, 0.44 ＋0.04; and the expressions of VEGF protein were 0. 14 ±0.02, 0. 12 ±0.01,0.95 ±0.09, 0.87 ±0.09, 0.55 ±0.06; and cell viability rates were 100%, （98.43 _＋2.88）%, （76.15 ± 0. 70 ） %, （53.87 ± 0.77 ） %, （35.23 ＋ 0.67 ） % ; while cell apoptotic rates were （5.2 -＋ 1.12） %, （5.74 ± 1.07）%, （6.82 ±1.85）%, （12.09 ±3.53）%, （31.88 ±6.95）% ; the cell migration distance of PANC1 cells were （43.24 ±3.67）%, （59.46 ±5.39）%, （80.56 ±8.05）%, （63.89 ±5.96）%, （9.09 ± 1.59 ）%. Compared with those of control group, the expressions of VEGF mRNA, VEGF and HIF-lct protein, cell migration distance showed a two-way variation （ ascending first and descending later） （ P 〈 0.05 ）, and the expression of HIF-lot mRNA and cell proliferation rate was decreased in a dose-dependent manner, while the cell apoptosis was increased in a dose-dependent manner. Conclusions CoCl2 significantly inhibits the proliferation and promotes apoptosis of PANC1 cells at certain level. CoCl2 has a two-way effect

关 键 词：胰腺肿瘤 缺氧 氯化钴 细胞增殖 细胞凋亡 细胞运动 

分 类 号：R735.9[医药卫生—肿瘤]