灰黄霉素对人前列腺癌PC-3细胞体内外作用的实验研究  

Inhibitory effect of griseofuivin on human prostate cancer PC-3 cells and Balb/c nude mice tumor xenograft

作　　者：段跃[1] 徐庆康[1] 于田强[1] 洪翔[1] 

机构地区：[1]武警浙江省总队医院泌尿外科,嘉兴314000

出　　处：《中华泌尿外科杂志》2012年第6期464-467,共4页Chinese Journal of Urology

摘　　要：目的探讨灰黄霉素在体内外对人前列腺癌PC-3细胞抗肿瘤作用的可能机制。方法不同浓度（0、5、10、15、20、25Izg／m1）的灰黄霉素作用于PC-3细胞48h，四甲基偶氮唑盐比色法测定细胞活率；荧光显微镜下观察细胞形态变化；膜联蛋白5一异硫氰酸荧光素／碘化丙啶双标记染色法检测PC-3细胞的凋亡；比色法检测Caspase．3、8、9活性变化。采用灰黄霉素对PC-3细胞BALB／C荷瘤裸鼠瘤体内注射治疗，观察肿瘤生长变化，12d后免疫组化法检测Bcl-2、Bax、p53和Survivin的表达。结果灰黄霉素能抑制PC-3细胞增殖，半数抑制率浓度为（18．17±2．10）μg／ml；镜下可见典型的细胞凋亡形态学改变。15μg／ml灰黄霉素处理组PC-3细胞凋亡率（31．37±2．93）％，与对照组（2．89±0．67）％比较差异有统计学意义（P〈0．01）。15μg／ml灰黄霉素处理组PC-3细胞caspase-3、8、9的活性分别为0．562±0．050、0．337±0．053和0．293±0．038，对照组分别为0．113±0．014、0．120±0．017和0．109±0．018，组间比较差异有统计学意义（P〈0．01）。肿瘤接种后第23天，灰黄霉素组移植瘤体积为（961．17±78．12）mm^2，移植瘤质量为（742．50±78．63）mg，对照组为（1732．96±120．73）mm^2，（1387．33±71．47）mg，组间比较差异有统计学意义（P〈0．01）。灰黄霉素组Bcl-2、Bax、p53和Survivin蛋白表达阳性率分别为（16．10±3．45）％、（39．50±6．88）％、（48．20±8．04）％和（16．50±2．22）％，对照组分别为（41．30±3．95）％、（13．70±2．98）％、（17．60±3．21）％和（52．11±6．28）％，组间比较差异有统计学意义（P〈0．01）。结论灰黄霉素能抑制前列腺癌PC-3细胞的生长，诱导PC-3细胞凋亡并抑制PC-3细胞的动物体内致瘤能力。Objective To study the anti-tumor effects of griseofulvin on human prostate cancer PC- 3 cells both in vitro and in vivo. Methods PC-3 cells were treated with griseofulvin at various concentrations for 48 h, cell survival rate was then measured by MTT assay. The changes of morphology were observed by fluorescence microscope; Annexin V-FITC apoptosis detection kit was used to detect apoptosis of the cells ; The enzyme activity changes of caspase-3, 8, 9 were detected by spectrophotometry. For in vivo study, we first established the PC-3 tumor model by grafting PC-3 cells in athymic nude mice, and then injected griseofulvin into the tumors. 12 days after injection, the mice were sacrificed, the tumors were removed, weighed and the ratios of tumor-suppression were then calculated. We had detected the expressions of Bcl-2, Bax, p53 and Survivin with immumohistochemistry as well. Results MTT results showed that griseofulvin could significantly inhibit the proliferation of PC-3 cells in vitro in a dose-dependent manner,and the ICS0 of griseofulvin was 18.17 ± 2.10 p,g/ml. Typical morphological changes of PC-3 cells were observed by microscope. The rates of apoptosis of griseofulvin treated PC-3 cells greatly increased compared with that of the control cells （31.37 ± 2.93 % vs 2.89 ± 0.67% , P 〈 0.01 ）. The caspase-3, caspase-8 and caspase-9 ac-tivities in griseofulvin treated PC-3 cells were significantly higher than those in control cells （ 0. 562 ± 0. 050 vs 0. 113 ±0.014, 0.337 ±0.053 vs 0. 120 ±0.017, 0.293 ±0.038 vs 0. 109 ±0.018, P 〈0.01）. On the 23th day after tumor vaccination, the tumor volume was 961. 17 ± 78. 12 mm^3 in griseofulvin treated group and was 433.6 ± 12.8 mm^3 in control group （P 〈0.01 ）. The tumor weight was 742.50 ± 78.63 mg in griseofulvin treated group and was 1387.33 ± 71.47 mg^3 in control group （P 〈 0. 01 ）. Bcl-2, Bax, p53 and Survivin protein expressions were 16.10± 3.45% , 39.50 ± 6.88% , 48.20 ± 8.04% , 16.50 ± 2.22% in griseofulvin treated grou

关 键 词：灰黄霉素 PC-3细胞 凋亡 体内成瘤 

分 类 号：R737.25[医药卫生—肿瘤]