表没食子儿茶素没食子酸酯抑制前列腺癌细胞PC-3组织蛋白酶B合成与分泌对其侵袭能力的影响  

Expression and secretion of cathepsin B involved in the inhibition of PC-3 cell invasion by epigallocatechin gallate

作　　者：滕若冰[1] 白璐 徐方明[1] 张森 卢亚乐 段杰 米超 高漓[1] Teng Ruobing;Bai Lu;Xu Fangming;Zhang Sen;Lu Yale;Duan Jie;Mi Chao;Gao Li(Department of Urology,Affiliated Hospital of Guilin Medical College,Guilin 541001,China)

机构地区：[1]桂林医学院附属医院泌尿外科,541001

出　　处：《中华实验外科杂志》2020年第3期489-492,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金项目(81660425);桂林市科学研究与技术开发项目(20170226)。

摘　　要：目的探讨表没食子儿茶素没食子酸酯(EGCG)对前列腺癌细胞PC-3侵袭的影响以及组织蛋白酶B(Cathepsin B)在其中可能的作用机制。方法使用不同浓度EGCG培养组织来源为人前列腺癌分离自骨转移灶的PC-3细胞(购自中国科学院上海细胞库),形态学检查分析PC-3细胞在侵袭、迁移上的改变;实验分为对照组与EGCG干预组(12.5、25.0、50.0、100.0 mg/L),蛋白质印迹法(Western blot),检测不同浓度EGCG对PC-3细胞Cathepsin B合成和分泌的影响;酶联免疫吸附(ELISA)实验,检测不同浓度的EGCG对PC-3细胞CathepsinB分泌的影响。研究历时2年。组间数据分析采用One-way ANOVA分析,组间两两比较采用Tukey方法。结果抑制侵袭实验中,除12.5 mg/L EGCG干预组(341.67±25.17,t=2.230,P>0.05)侵袭细胞数相比对照组(376.33±9.50)差异无统计学意义外,25.0 mg/L EGCG干预组(241.00±3.60,t=23.070,P<0.05)、50.0 mg/L EGCG干预组(170.67±13.31,t=21.780,P<0.05)和100.0 mg/L EGCG干预组(72.66±6.80,t=45.020,P<0.05)相比于对照组明显减少,差异均有统计学意义(F=241.050,P<0.01);抑制PC-3迁移实验中,12.5 mg/L EGCG干预组侵袭细胞数(0.550±0.030,t=5.730,P<0.05)、25.0 mg/L EGCG干预组(0.430±0.018,t=20.740,P<0.05)、50.0 mg/L EGCG干预组(0.320±0.025,t=22.620,P<0.05)和100.0 mg/L EGCG干预组(0.260±0.017,t=38.830,P<0.05)相比对照组(0.650±0.004)明显减少,差异均有统计学意义(F=179.60,P<0.01);Western blot结果显示,除12.5 mg/L EGCG干预组(3.870±0.151,t=0.710,P>0.05)CathepsinB蛋白的合成和分泌相比对照组(3.99±0.25)差异无统计学意义外,25.0 mg/L EGCG干预组(3.29±0.12,t=4.370,P<0.05)、50.0 mg/L EGCG干预组(2.88±0.10,t=7.140,P<0.05)和100.0 mg/L EGCG干预组(2.060±0.085,t=12.660,P<0.05)相比于对照组明显减少,差异均有统计学意义(F=79.080,P<0.01)。ELISA实验显示,除12.5 mg/L EGCG干预组上清的Cathepsin B前酶光密度值比值(3.00±0.17,t=2.040,P>0.05)和总组织蛋白酶B含量(50.23±1.75,t=Objective To study the effects of epigallocatechin gallate(EGCG)on invasion of human prostatic carcinoma cell PC-3,and explore the potential mechanism of CathepsinB involved in the process.Methods Different concentrations of EGCG were used to culture PC-3 cells derived from bone metastases from human prostate cancer,and morphological examination was performed to analyze the changes of PC-3 cells’invasion,migration and other metastatic capabilities;through immunoblotting experiments,the detection of different concentrations of EGCG The effect of Cathepsin B protein synthesis in PC-3 cells;Western blot and ELISA experiments were used to detect the effect of different concentrations of EGCG on the secretion of Cathepsin B protein in PC-3 cells.The study period is 2 years.Analysis using SPSS 26.0 statistical software.One-way analysis of variance was used for data analysis between groups,and Tukey method was used for comparison between groups.P<0.05 was considered statistically significant.Results In the experiment of inhibiting invasion,except 12.5 mg/L was no significant difference in the number of invasive cells between the two groups(341.67±25.17,t=2.230,P>0.05).In addition,25.0 mg/L group(241.00±3.60,t=23.070,P<0.05),50.0 mg/L group(170.67±13.31,t=21.780,P<0.05),and 100.0 mg/L group(72.66±6.80,t=45.020,P<0.05),the number of invasive cells were significantly reduced compared with the control group(376.33±9.50),and the differences between the groups were statistically significant(F=241.050,P<0.01);In the inhibition of PC-3 migration experiment,the number of invasive cells in 12.5 mg/L EGCG group(0.550±0.030,t=5.730,P<0.05),25.0 mg/L EGCG group(0.430±0.018,t=20.740,P<0.05),50.0 mg/L EGCG group(0.320±0.025,t=22.620,P<0.05)and 100.0 mg/L EGCG group(0.260±0.017,t=38.830,P<0.05)was significantly reduced compared with the control group(0.650±0.004),and the differences between the groups were statistically significant(F=179.60,P<0.01);Western blotting results showed that except 12.5 mg/L(3.870±0.151,t=0.710,P

关 键 词：表没食子儿茶素没食子酸酯 前列腺癌 侵袭 组织蛋白酶B 

分 类 号：R737.25[医药卫生—肿瘤]