壁虎活性多肽对线粒体通路诱导的人食管癌Ec 9706细胞增殖与凋亡的影响  

Effect of gecko active polypeptide on the proliferation and apoptosis of human esophageal carcinoma 9706 cells inducing by mitochondria pathway

作　　者：吕行直 李瑞芳[1] 李钟杰 刘玲[1] 王建刚[1] LV Xing-zhi;LI Rui-fang;LI Zhong-jie;LIU Ling;WANG Jian-gang(The Key Laboratory of Pharmacologyand Medical Molecular Biology, MedicalCollege, Henan University of Science andTechnology, Luoyan^ 471023, HenanProiirtcc, China)

机构地区：[1]河南科技大学医学院药理学与分子生物学重点实验室,河南洛阳471023

出　　处：《中国临床药理学杂志》2019年第6期543-546,共4页The Chinese Journal of Clinical Pharmacology

基　　金：河南省科技攻关重点基金资助项目(142102310031)

摘　　要：目的研究壁虎活性多肽(GAP)对人食管癌Ec 9706细胞的增殖与凋亡的影响。方法将7个不同质量浓度(0. 3,1,3,10,30,100和300μg·m L^(-1))的GAP处理Ec9706细胞48 h,以CCK8法检测细胞增殖水平,根据CCK8的结果,将细胞分为4组:对照组(不加药)和低、中、高3个剂量(GAP:15,30,60μg·m L^(-1))实验组。用流式细胞仪检测Ec 9706细胞的凋亡率;用蛋白免疫印迹法检测细胞线粒体凋亡通路B淋巴细胞瘤-2基因(Bcl-2)、Bcl-2相关X蛋白(Bax)和胱天蛋白酶3(Caspase3)蛋白的表达情况。结果 GAP在24,48 h的IC50分别为70. 2和54. 6μg·m L^(-1)。处理48 h后,对照组和低、中、高3个剂量实验组凋亡率分别为(6. 24±1. 73)%,(35. 40±6. 33)%,(50. 60±8. 61)%和(84. 70±8. 89)%,3个剂量实验组与对照组比较,差异均有统计学意义(均P <0. 01)。这4组的Bax蛋白相对灰度值分别为0. 55±0. 07,0. 61±0. 06,0. 77±0. 11和1. 07±0. 14;这4组的Bcl-2蛋白相对灰度值分别为0. 80±0. 13,0. 54±0. 11,0. 36±0. 04和0. 11±0. 03;这4组的Caspase3蛋白相对灰度值分别为0. 45±0. 05,0. 51±0. 09,0. 92±0. 12和0. 98±0. 16。3个剂量实验组与对照组比较,以上各指标的差异均有统计学意义(P <0. 05或P <0. 01)。结论 GAP可以显著抑制食管癌Ec 9706细胞增殖,并且诱导细胞凋亡,其机制与其调控线粒体凋亡通路相关蛋白表达有关。Objective To investigate the effects of gecko active polypeptide( GAP) on the proliferation and apoptosis of Ec 9706 cells.Methods Ec9706 cells were treated with seven different concentrations( 0.3,1,3,10,30,100 and 300μg·m L^-1) of GAP for 48 h.The CCK8 assay was used to assess cell proliferation.According to CCK8 results,the cells were divided into four groups: control group( untreated),experimental-L,experimental-M and experimental-H groups( 15,30,60μg·m L^-1 GAP).Cell apoptosis was observed by flow cytometry.The expressions of mitochondrial apoptosis pathway marker protein B-cell lymphoma( Bcl-2),Bcl-2 associated X protein( Bax),Cysteinyl aspartate specific proteinase 3( Caspase 3) were detected by Western-blotting.Results After treatment of GAP for 24 h and 48 h,the 50%inhibitory dose( IC50) values were 70.2 μg·m L^-1 and 54.6μg·m L^-1.After administration for 48 h,the apoptotic rates in control group,and experimental-L,experimental-M and experimental-H groups were( 6.24±1.73)%,( 35.40±6.33)%,( 50.60±8.61)% and( 84.70±8.89)%;comparison between three doses experimental groups with the control group,the difference was statistically significant( all P < 0.01).The protein levels of Bax in the 4 groups were0.55±0.07,0.61±0.06,0.77±0.11,and 1.07±0.14,respectively;the Bcl-2 protein expressions in the 4 groups were 0.80±0.13,0.54±0.11,0.36±0.04 and 0.11±0.03,respectively;the protein expressions of Caspase3 in 4 groups were 0.45±0.05,0.51±0.09,0.92±0.12 and 0.98±0.16.Comparison between three doses experimental groups with the control group,the difference of the three factors were statistically significant( P < 0.05 or P < 0.01).Conclusion GAP inhibited the proliferation and induced the apoptosis of Ec9706 cells via targeting the apoptotic mitochondrial pathway.

关 键 词：壁虎活性多肽 EC 9706细胞 细胞凋亡 线粒体通路 

分 类 号：R28[医药卫生—中药学]