CD147拮抗肽和基质金属蛋白酶抑制剂多西环素对胆囊癌细胞的作用  

Effect of CD147 antagonist peptide and doxycycline on gallbladder carcinoma cells

作　　者：王世行[1] 刘超[1] 黄涛 刘新江[1] 罗乾坤 庞志刚[1] 

机构地区：[1]郑州大学第二附属医院普外科,450014 [2]郑州大学附属肿瘤医院肝胆外科,450008

出　　处：《中华实验外科杂志》2017年第7期1145-1148,共4页Chinese Journal of Experimental Surgery

基　　金：河南省二0一五年科技发展计划（152102310256）

摘　　要：目的观察CD147拮抗肽-9（AP-9）及多西环素（DOX）分别及联合用药对胆囊癌细胞（GBC-SD）的增殖、凋亡、CD147表达和基质金属蛋白酶（MMP）-9活性的影响。方法将AP-9（50、100、200、400 μg/ml）、DOX（5、10、20、40 μg/ml）和联合用药（AP-9 200 μg/ml ＋ DOX 20 μg/ml）作用于胆囊癌细胞12、24、48 h，细胞计数试剂盒（CCK-8）法检测细胞增殖，Western blot法检测CD147表达，明胶酶谱法检测MMP-9活性，膜联蛋白V（Annexin V）/碘化丙锭（PI）染色法流式细胞仪检测早期凋亡率，结果采用t检验。结果不同浓度AP-9处理组的早期凋亡率（2.13±0.10）%、（2.92±0.19）%较对照组（1.58±0.08）%升高（P=0.002、P=0.001）；MMP-9活性（320.08±33.90、249.59±33.63、259.63±34.08、172.31±35.20）较对照组（446.08±42.42）降低（P=0.013、0.002、0.003、0.001）；增殖抑制率较对照组升高（P均=0.001）且呈时间和剂量依赖趋势。不同浓度多西环素处理组的早期凋亡率（3.25±0.12）%、（3.63±0.24）%较对照组（1.58±0.08）%明显升高（P均=0.001）；MMP-9活性（203.52±18.25、175.22±23.30、147.86±22.83、109.99±29.22）较对照组（446.08±42.42）降低（P均=0.001）；增殖抑制率较对照组升高（P均=0.001），CD147表达（0.97±0.12、0.80±0.10、0.69±0.11、0.51±0.12）较对照组（2.04±0.15）明显降低（P均=0.001）。联合用药后较单独用药，细胞增殖抑制率（38.78±0.39）%、（43.76±0.71）%、（52.00±0.91）%升高（P=0.001、P=0.003），MMP-9活性（110.19±24.74）降低（P=0.001、P=0.024），早期凋亡率（3.73±0.29）%升高（P=0.001、P=0.048）。结论DOX可减少胆囊癌细胞CD147表达，AP-9和DOX单独或联合用药均可以抑制胆囊癌细胞增殖、抑制MMP-9活性，促进细胞凋亡，且联合用药后抑制作用增强。Objective To research the effect of GBC-SD on proliferation, apoptosis, expression of CD147 and activity of matrix metalloproteinase （MMP）-9 in gallbladder carcinoma cells, with single and combination therapy of CD147 antagonist peptide and doxycycline.Methods Gallbladder carcinoma cells were treated with antagonist peptide （AP-9） （50, 100, 200, 400 μg/ml） or DOX （5, 10, 20, 40 μg/ml） alone and combination therapy （AP-9 200 μg/ml ＋ DOX 20 μg/ml） for 12, 24 and 48 h, then the inhibition rate of proliferation was detected by cell counting kit-8 （CCK-8）, expression of CD147 was detected by Western blotting, the activity of MMP-9 was detected by gelatin zymography, and the early apoptosis rate was detected by flow cytometry with Annexin V/propidium iodide （PI） staining.Results After treatment with several concentrations of AP-9, the apoptosis rate [（2.13±0.10）% and （2.92±0.19）%] was higher （P=0.002, P=0.001） than control group [（1.58±0.08）%]; the activity of MMP-9 [（320.08±33.90）, （249.59±33.63）, （259.63±34.08） and （172.31±35.20）] was reduced （P=0.013, 0.002, 0.003 and 0.001 respectively） as compared with that in control group; and cell proliferation was inhibited as compared with that in control group （P=0.001）. After treatment with several concentrations of doxycycline, the apoptosis rate [（3.25±0.12）% and （3.63±0.24）%] was higher （P=0.001） than in the control group [（1.58±0.08）%]; the activity of MMP-9 [（203.52±18.25）, （175.22±23.30）, （147.86±22.83） and （109.99±29.22）] was reduced （P=0.001） as compared with control group [（446.08±42.42）]; cell proliferation was inhibited as compared with control group （P=0.001）; the expression level of CD147 [（0.97±0.12）, （0.80±0.10）, （0.69±0.11） and （0.51±0.12）] was reduced （P=0.001） as compared with cotrol group [（2.04±0.15）]. As compared with treatment alone, cell proliferation [（38.78±0.39）%, （43.76±0.71）% a

关 键 词：胆囊癌 CD147 细胞外基质金属蛋白酶-9 拮抗肽 多西环素 

分 类 号：R735.8[医药卫生—肿瘤]