FK506结合蛋白5对人前列腺癌LNCaP细胞株及裸鼠移植瘤的影响  

The preliminary study of the effect of FK506 binding protein 5 expression on cell proliferation in human prostate cancer LNCaP cell line and xenograft in nude mice

作　　者：王强[1] 胡斌[2] 郝海峰[3] 尚芝群[1] 牛远杰[1] 权昌益[1] 

机构地区：[1]天津医科大学第二医院泌尿外科天津市泌尿外科研究所,300211 [2]天津医科大学总医院泌尿外科 [3]山西医科大学太钢总医院泌尿外科

出　　处：《中华实验外科杂志》2015年第12期2932-2935,共4页Chinese Journal of Experimental Surgery

基　　金：天津市卫生局科技基金资助项目（2012KZ090）;国家自然科学基金资助项目（81302211）;国家重大基础研究计划（973）资助项目（2012CB518304）

摘　　要：目的观察抑制FK506结合蛋白5（FKBP5）基因表达对前列腺癌LNCaP细胞株及裸鼠移植肿瘤生长的影响。方法利用特异性针对FKBP5的短发夹RNA（shRNA）和FKBP5抑制剂他克莫司（FK506）处理前列腺癌LNCaP细胞，然后用噻唑蓝比色法（MTr）检测细胞生长情况，并通过Westernblot检测FK506处理过的LNCaP细胞中凋亡相关蛋白的变化；另外构建LNCaP细胞裸鼠皮下移植瘤模型，并用FK506给裸鼠灌胃，对比实验组及对照组皮下肿瘤生长情况。结果利用shRNA抑制LNCaP细胞中FKBP5的表达后，实验组细胞的增殖活性明显低于对照组。MTF法检测实验组的吸光度（A）值为0．359±0．021，而对照组为0．631±0．025（P〈0．05）。此外，FK506对细胞生长的抑制作用具有浓度依赖性，MTY法检测实验组（0．1、1．0、10．0μmol／L）A值分别为1．309±0．209、0．875±0．142、0．390±0．046，对照组为1．403±0．103（P〈0．05）。Westernblot检测天冬氨酸蛋白水解酶-3（Caspase-3）表达量亦随之增加；此外，利用FKS06抑制体内FKBP5的表达后，小鼠肿瘤生长受到明显抑制。非去势组中，处理组小鼠肿瘤体积为（123．53±6．56）mm3明显小于对照组[（220．88±6．44）mm2，P〈0．05]；去势组中，处理组小鼠肿瘤体积为（103．71±7．17）mm2，而对照组为[（194．07±4．93）mm2，P〈0．05]。结论FKBP5对于前列腺癌的生长起着重要作用。Objective To investigate roles of FKS06 binding protein 5 （ FKBP5 ） roles for the growth of prostate cancer cells and xenograft in nude mice. Methods LNCaP cells were treated with shRNA specifically for FKBP5 and FKS06 （ FKBP5 inhibitor）, then we detected the growth activity of these cells by methyl thiazol tetrazolium （MTY） assay; Besides, apoptosis proteins were detected by west- ern blot. Meanwhile, subcutaneous xenografts in mice model were established with LNCaP cells, then we compared the growth rates between FKS06 treated group and control group. Results The growth rate of the prostate cancer LNCaP cells was decreased significantly when FKBP5 was knockdown by shRNA. The opti- cal density of the experimental group（0. 359 ＋0. 021 ）detected by MTr was significantly lower than that of the control group（0. 631± 0. 025, P 〈 0. 05）. Besides, cell growth was suppressed in a dose dependent manner by FKS06. The optical density of the experimental group （0. 1, 1.0, 10. 0 I∽mol/L） was 1. 309 ± 0. 209, 0. 875±0. 142, 0. 390 ±0. 046, while the control group was （ 1. 403±0. 103, P 〈 0. 05 ）. Expression of cysteinyl aspartate specific proteinase 3 （ Caspase - 3 ） was higer with the increasing concen- tration of FK506. In mice model, tumors were significantly inhibited when treated with FK506. Tumor si- zes in experimental group were smaller [ （ 123.53 ± 6. 56） mm3 vs. （220. 88± 6.44） mm3 P 〈 0. 05 ] The same results were obtained even if mice were initially castrated before [ （103.71± 7.17 ） mm3 vs. （ 194. 07±4. 93 ） mm3 , P 〈 0. 05 ]. Conclusion These results suggested that FKBP5 played important roles in the growth of prostate cancer, and might be one potential therapeutic target of prostate cancer.

关 键 词：前列腺癌 FK506结合蛋白5 他克莫司 天冬氨酸蛋白水解酶-3 

分 类 号：R979.5[医药卫生—药品]