So Alan, Sinnemann Shannon, Huntsman David, Fazli Ladan, Gleave Martin
The Prostate Centre, Vancouver General Hospital, University of British Columbia, Vancouver, British Columbia, Canada.
Mol Cancer Ther. 2005 Dec;4(12):1837-49. doi: 10.1158/1535-7163.MCT-05-0178.
Clusterin is a stress-associated cytoprotective chaperone up-regulated by various apoptotic triggers in many cancers and confers treatment resistance when overexpressed. The objectives of this study were to evaluate clusterin expression levels in human breast cancer and to determine whether antisense oligonucleotides or double-stranded small interfering RNAs (siRNA) targeting the clusterin gene enhance apoptosis induced by paclitaxel. Clusterin immunostaining was evaluated in a tissue microarray of 379 spotted breast cancers. The effect of hormone withdrawal, paclitaxel treatment, clusterin antisense oligonucleotide (OGX-011), and siRNA treatments on clusterin expression was examined in MCF-7 and MDA-MB-231 cells. Northern, quantitative real-time PCR, and Western analyses were used to measure change in clusterin mRNA and protein levels. The effect of OGX-011 or siRNA clusterin treatment on chemosensitivity to paclitaxel was done in both cell lines in vitro, whereas the ability of OGX-011 to chemosensitize in vivo was evaluated in athymic mice bearing MCF-7 tumors. Clusterin was expressed in 62.5% of tumors within the tissue microarray. Clusterin expression increased after estrogen withdrawal and paclitaxel treatment in vitro in MCF-7 cells. OGX-011 or siRNA clusterin decreased clusterin levels by >90% in a dose-dependent, sequence-specific manner and significantly enhanced chemosensitivity to paclitaxel in vitro. When combined, OGX-011 or siRNA clusterin reduced the IC50 by 2-log compared with paclitaxel alone. In vivo administration of OGX-011 enhanced the effects of paclitaxel to significantly delay MCF-7 tumor growth. These data identify clusterin as a valid therapeutic target and provides preclinical proof-of-principle to test OGX-011 in multimodality therapies for breast cancer.
簇集素是一种与应激相关的细胞保护伴侣蛋白,在许多癌症中被各种凋亡触发因素上调,过表达时会赋予治疗抗性。本研究的目的是评估人乳腺癌中簇集素的表达水平,并确定靶向簇集素基因的反义寡核苷酸或双链小干扰RNA(siRNA)是否能增强紫杉醇诱导的细胞凋亡。在一个包含379个斑点状乳腺癌的组织芯片中评估了簇集素免疫染色。在MCF-7和MDA-MB-231细胞中检测了激素撤除、紫杉醇治疗、簇集素反义寡核苷酸(OGX-011)和siRNA处理对簇集素表达的影响。采用Northern印迹、定量实时PCR和Western分析来测量簇集素mRNA和蛋白水平的变化。在两种细胞系中体外研究了OGX-011或siRNA簇集素处理对紫杉醇化疗敏感性的影响,而在携带MCF-7肿瘤的无胸腺小鼠中评估了OGX-011体内化疗增敏能力。在组织芯片中的62.5%的肿瘤中表达了簇集素。在体外,雌激素撤除和紫杉醇治疗后,MCF-7细胞中簇集素表达增加。OGX-011或siRNA簇集素以剂量依赖性、序列特异性方式使簇集素水平降低>90%,并在体外显著增强了对紫杉醇的化疗敏感性。联合使用时,与单独使用紫杉醇相比,OGX-011或siRNA簇集素使IC50降低了2个对数。OGX-011的体内给药增强了紫杉醇的作用,显著延迟了MCF-7肿瘤的生长。这些数据确定簇集素是一个有效的治疗靶点,并为在乳腺癌多模式治疗中测试OGX-011提供了临床前原理证明。