钙调蛋白依赖性蛋白激酶Ⅱ对前列腺癌PC3细胞增殖、侵袭及上皮-间充质转化的影响  

Effects of calcium/calmodulin dependent protein kinase Ⅱ on proliferation, invasion and epithelia - mesenchymai transition of prostate cancer PC3 cells

作　　者：彭璇[1] 陈晖[1] 王敏[1] 刘修恒[1] 

机构地区：[1]武汉大学,人民医院泌尿外科,430060

出　　处：《中华实验外科杂志》2015年第12期2957-2960,共4页Chinese Journal of Experimental Surgery

摘　　要：目的观察钙调蛋白依赖性蛋白激酶Ⅱ（CaMKⅡ）活化的抑制对人前列腺癌PC3细胞增殖、侵袭能力及上皮一间充质转化（EMT）相关信号通路的影响。方法通过CaMKⅡ特异性抑制剂KN93处理PC3细胞，抑制其活化后，采用噻唑蓝（Mrrr）法检测PC3细胞增殖抑制率，Transwell小室法检测PC3细胞侵袭能力，Westernblot测定PC3细胞中磷酸化CaMKII（p-CaMKII）、核因子-KB（NF-KB）、锌指转录因子（Snail）、Raf激酶抑制蛋白（RKIP）的表达。结果不同浓度KN93处理PC3细胞24h后，5、10、20p,mol／L药物组p-CaMKII蛋白表达（0．453±0．070、0．368±0．076、0．308±0．011）较对照组（0．596±0．028）显著减少（P〈0．05），40g,mol／L药物组未见明显蛋白表达。各药物组24h增殖抑制率分别为（6．88±1．79）％、（12．92±2．74）％、（17．88±2．86）％、（31．23±4．24）％；48h增殖抑制分别为（16．53±2．45）％、（29．02±1．74）％、（40．52±1．98）％、（52．26±3．51）％。Transwell小室培养48h后，对照组和各药物组穿膜细胞数分别为（149±17）、（97±7）、（59±9）、（51±7）、（24±3）μmol／高倍视野，各药物组穿膜细胞数均明显减少（P〈0．01）。Westernblot结果示，对照组和各药物组NF．KBp65蛋白的相对表达量差异无统计学意义（P〉0．05）。而各药物组p-NF-KBp65蛋白的相对表达量（0．483±0．052、0．490±0．064、0．432±0．057、0．341±0．008）较对照组（0．597±0．020）明显降低（P〈0．05）。与对照组（0．716±0．046）比较，5、10μmol／L药物组Snail蛋白的相对表达量（0．337±0．058、0．137±0．045）显著降低（P〈0．01），20、40μmol／L药物组甚至未见明显蛋白表达。而药物组RKIP蛋白的相对表达量（0．720±0．003、0．732±0．008、0．991±0．025、1．116±0．020）显著高于对照组（0．372±0．010，P〈0．01）。结论CaMKObjective To observe the effects of inactivation of calcium/calmodulin dependent pro- tein kinase Ⅱ （ CaMK Ⅱ ） on proliferation, invasion and epithelia - mesenchymal transition related signa- ling pathway in prostate cancer PC3 cells. Methods The activity of CaMK II in PC3 cells was suppressed by KN93, a pharmacological inhibitor. MTT assay was used to detect the inhibition rate of PC3 cells and the invasion ability of PC3 cells was examined using Transwell invasion chambers. The protein expression of phosphorylated CaMKⅡ （p -CaMKⅡ ）, nuclear factor KB （NF - KB）, zinc finger transcription factor （Snail）, and Raf kinase inhibitory protein （RKIP） was measured by Western blotting. Results After treatment with KN93 for 24 h, the protein expression of p - CaMK Ⅱ PC3 cells treated with 5, 10, 20μmol/L KN93 （0. 453 ± 0. 070, 0. 368 ± 0. 076, and 0. 308 ± 0. 011 ） was significantly decreased as compare with control group （ 0. 596 ± 0. 028 ） （ P 〈 0.05 or 0.01 ）, 40 ixmol/L KN93 almost completely inhibited p - CaMK I1 protein expression. The inhibition rate of PC3 cells treated with KN93 for 24 h was （6.88±1.79）%, （12.92 ±2.74）%, （17.88 ±2.86）% and （31.23±4.24）%, and （16.53 ± 2.45）%, （29.02±1.74）%, （40.52 ±1.98）% and （52.26± 23.51）% for 48 h respectively. The number of the invasion PC3 cells in control and KN93 groups was （ 149 ±17 ）, （ 97± 7 ）, （ 59 ±9 ）, （51 ±7）, and （24 ± 3）/high magnification mirror （HP） respectively. The number of invasion cells in KN93 groups was significantly decreased when compared with control group （ P 〈 0. 01 ）. The protein ex- pression of NF - KB p65 had no significant difference between control and KN93 groups （ P 〉 0.05 ） , but p - NF - KB p65 was down - regulated in KN93 groups （ 0. 483 ± 0. 052, 0. 490 ±0. 064, 0. 432 ± 0. 057, and 0. 341 ±0. 008） when compared with control group （0. 597 ±0. 020, P 〈0. 05 or 0. 01 ）. As compared with control group

关 键 词：前列腺癌 钙调蛋白依赖性蛋白激酶Ⅱ 增殖 侵袭 上皮-间充质转化 

分 类 号：R284.2[医药卫生—中药学]