1,25D3 增强了吉西他滨和顺铂在人膀胱癌模型中的抗肿瘤活性。
1,25D3 enhances antitumor activity of gemcitabine and cisplatin in human bladder cancer models.
机构信息
Department of Pharmacology and Therapeutics, Roswell Park Cancer Institute, Buffalo, New York, USA.
出版信息
Cancer. 2010 Jul 1;116(13):3294-303. doi: 10.1002/cncr.25059.
BACKGROUND
1,25 dihydroxyvitamin D3 (1,25D3) potentiates the cytotoxic effects of several common chemotherapeutic agents. The combination of gemcitabine and cisplatin is a current standard chemotherapy regimen for bladder cancer. The authors investigated whether 1,25D3 could enhance the antitumor activity of gemcitabine and cisplatin in bladder cancer model systems.
METHODS
Human bladder cancer T24 and UMUC3 cells were pretreated with 1,25D3 followed by gemcitabine and cisplatin. Apoptosis was assessed by annexin V staining. Caspase activation was examined by immunoblot analysis and substrate-based caspase activity assay. The cytotoxic effects were examined by using 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) and in vitro clonogenic assay. p73 protein levels were assessed by immunoblot analysis. Knockdown of p73 was achieved by siRNA. The in vivo antitumor activity was assessed by in vivo excision clonogenic assay and tumor regrowth delay in the T24 xenograft model.
RESULTS
1,25D3 pretreatment enhanced gemcitabine and cisplatin-induced apoptosis and the activities of caspases 8, 9, and 3 in T24 and UMUC3 cells. 1,25D3 synergistically reduced gemcitabine and cisplatin-suppressed surviving fraction in T24 cells. 1,25D3, gemcitabine, or cisplatin induced p73 accumulation, which was enhanced by gemcitabine and cisplatin or 1,25D3 and gemcitabine and cisplatin. p73 expression was lower in human primary bladder tumor tissue compared with adjacent normal tissue. Knockdown of p73 increased clonogenic capacity of T24 cells treated with 1,25D3, gemcitabine and cisplatin, or 1,25D3 and gemcitabine and cisplatin. 1,25D3 and gemcitabine and cisplatin combination enhanced tumor regression compared with 1,25D3 or gemcitabine and cisplatin alone.
CONCLUSIONS
1,25D3 potentiates gemcitabine and cisplatin-mediated growth inhibition in human bladder cancer models in vitro and in vivo, which involves p73 induction and apoptosis.
背景
1,25 二羟维生素 D3（1,25D3）可增强几种常见化疗药物的细胞毒性作用。吉西他滨和顺铂联合是当前膀胱癌的标准化疗方案。作者研究了 1,25D3 是否可以增强膀胱癌模型系统中吉西他滨和顺铂的抗肿瘤活性。
方法
人膀胱癌 T24 和 UMUC3 细胞用 1,25D3 预处理，然后用吉西他滨和顺铂处理。用 Annexin V 染色评估细胞凋亡。通过免疫印迹分析和基于底物的半胱天冬酶活性测定来检查半胱天冬酶的激活。通过 3-(4,5-二甲基-2-噻唑基)-2,5-二苯基-2H-四唑溴盐（MTT）和体外集落形成试验来检测细胞毒性作用。通过免疫印迹分析评估 p73 蛋白水平。通过 siRNA 实现 p73 的敲低。通过体内切除集落形成测定和 T24 异种移植模型中的肿瘤再生长延迟来评估体内抗肿瘤活性。
结果
1,25D3 预处理增强了 T24 和 UMUC3 细胞中吉西他滨和顺铂诱导的细胞凋亡和半胱天冬酶 8、9 和 3 的活性。1,25D3 协同降低了 T24 细胞中吉西他滨和顺铂抑制的存活分数。1,25D3、吉西他滨或顺铂诱导 p73 积累，吉西他滨和顺铂或 1,25D3 和吉西他滨和顺铂增强了 p73 积累。与相邻正常组织相比，人原发性膀胱癌组织中的 p73 表达水平较低。p73 表达的敲低增加了用 1,25D3、吉西他滨和顺铂或 1,25D3 和吉西他滨和顺铂处理的 T24 细胞的集落形成能力。与单独使用 1,25D3 或吉西他滨和顺铂相比，1,25D3 和吉西他滨和顺铂联合增强了肿瘤消退。
结论
1,25D3 增强了人膀胱癌模型中吉西他滨和顺铂介导的体外和体内生长抑制，这涉及 p73 诱导和细胞凋亡。
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