斑蝥素和去甲斑蝥素抑制膀胱肿瘤细胞增殖的实验研究  

Study on the antineoplastic effect of cantharidin and norcantharidin for carcinoma cell of bladder

作　　者：沈海山 宋志强 李胜文 吴建臣 张卫东 李强 李世海 聂义鑫 

机构地区：[1]清华大学第一附属医院泌尿外科,北京100016

出　　处：《中华临床医师杂志（电子版）》2015年第3期86-90,共5页Chinese Journal of Clinicians(Electronic Edition)

基　　金：清华-周大福医学研究专项基金(202836019-04);清华-裕元医学科学研究基金(20240000538)

摘　　要：目的通过对比研究不同浓度斑蝥素（CA）和去甲斑蝥素（NCTD）对体外人膀胱肿瘤EJ、T24和鳞状细胞癌细胞（BSCC）增殖的抑制作用，明确药物效果、最佳药物浓度其抗肿瘤细胞增殖的机制。方法使用不同浓度的CA和NCTD与肿瘤细胞共同培养，MTT实验计算细胞生长抑制率，进而选出抑制肿瘤细胞的有效药物及其最佳浓度。选用最佳浓度的最佳药物与肿瘤细胞共同培养，不同时间收集细胞，电子显微镜及激光共聚焦显微镜观察细胞形态。结果CA在体外对人膀胱肿瘤EJ、T24和BSCC细胞增殖有明显抑制作用，NCTD对于EJ和T24细胞有增殖有明显抑制作用，对BSCC细胞增殖的无明显抑制作用。在浓度小于40μmol／L、作用时间12-24h的情况下，CA对EJ和T24细胞的增殖抑制作用具有明显的时间-效应和剂量．效应关系，24h时药物半数抑制浓度（IC50）为20μmol／L；在浓度小于60μmol／L、作用时间24-48h情况下，CA对BSCC的抑制作用具有明显的时间．效应和剂量．效应关系，48h时药物IC50为60μmol／L。在浓度小于80gmol／L、作用时间24-48h情况下，NCTD对EJ和T24细胞的增殖抑制作用具有明显的时间．效应和剂量．效应关系；24h时药物IC50为60μmol／L。激光共聚焦扫描显微镜及电子显微镜观察证实CA和NCTD通过诱导膀胱肿瘤细胞凋亡进而抑制其增殖。结论CA及NCTD均具有较好的抑制常见膀胱肿细胞增殖的作用，有望用于浅表膀胱肿瘤电切术后的膀胱灌注化疗；且CA对BSCC细胞增殖具有较好的抑制作用，有望用于膀胱肿瘤电切标本中伴发BSCC患者的膀胱灌注化疗。Objective Cantharidin （CA） is recently used as anti-tumor natural medicine in many cancers including bladder carcinoma, but the mechanisms are poorly understood. This study is aimed to explore the inhibit effects of CA and its derivative-norcantharidin （NCTD） on T24, EJ and squarnous carcinoma cell （BSCC） of bladder in vitro. Methods Cultured carcinoma cells were treated with CA or NCTD at different concentrations for 12 h, 24 h, 36 h, 48 h and 60 h, then cell viability was measured by MTT assay. After treated by CA and NCTD at optimized concentrations, careinorna cells cellular morphological changes were also investigated by using Laser Scanning Confoeal Microscope （LSCM） and Electron Microscope. Results Human bladder center cell proliferation of EJ, T24 and BSCC cells significantly was inhibited in vitro by CA. EJ and T24 cell proliferation obviously was inhibited by NCTD, and BSCC cell can not be inhibited by NCTD. In a concentration of less than 40 μmol/L, between 12-24 h, the inhibitory effect of CA on proliferation of EJ and T24 cells showed time-effect and dose-effect relationship, drug half inhibitory concentration （IC50） of 24 h is 20μmol/L. In a concentration of less than 60μmol/L, between 24-48 h, the inhibition effect of CA on BSCC showed time-effect and dose-effect obvious relationship, IC50 of 48 h was 0μmol/L. In a concentration of less than 80μmol/L, between 24-48 h,inhibitory effect of NCTD on proliferation of EJ and T24 cells showed time-effect and dose-effect relationship; IC50 of 24 h was 60 μmol/L. LSCM and electron microscopy confirmed that CA and NCTD inhibited the proliferation of bladder cancer cells through the inducting apoptosis. Both CA and NCTD were able to decrease BSCC viability in time dependent and dose dependent manners; the maximum effect dose was 60μmol/L. When ceils were treated with CA or NCTD at 60 pmol/L for 24 h or 36 h, apoptosis was induced in BSCC, more Apoptotic bodies were founded in CA group. Conclusion CA and NCTD can inhibit the prolifer

关 键 词：斑蝥素 膀胱 癌 鳞状细胞 细胞增殖 去甲斑蝥素 

分 类 号：R737.14[医药卫生—肿瘤]