Knockdown of the cytoprotective chaperone, clusterin, chemosensitizes human breast cancer cells both in vitro and in vivo
Knockdown of the cytoprotective chaperone, clusterin, chemosensitizes human breast cancer cells both in vitro and in vivo
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DOI：
10.1158/1535-7163.mct-05-0178
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发表时间：
2005-12-01
影响因子：
5.7
通讯作者：
Gleave, M
中科院分区：
文献类型：
--
作者：
So, A;Sinnemann, S;Gleave, M
Clusterin是一种应激相关的细胞保护性伴侣蛋白，在许多癌症中由各种凋亡触发因子上调，当过度表达时会产生治疗耐药性。本研究的目的是评估聚集素在人乳腺癌中的表达水平，并确定针对聚集素基因的反义寡核苷酸或双链小干扰RNA(SiRNA)是否增强紫杉醇诱导的细胞凋亡。在379个点状乳腺癌的组织微阵列中对聚集素免疫染色进行了评估。观察激素停用、紫杉醇处理、聚集素反义寡核苷酸(OGX-011)和siRNA处理对MCF-7和MDA-MB-231细胞聚集素表达的影响。Northern、实时定量聚合酶链式反应和Western分析被用来测量聚集素mRNA和蛋白水平的变化。体外实验观察OGX-011或siRNA Clusterin对紫杉醇化疗敏感性的影响，在裸鼠MCF-7肿瘤模型上观察OGX-011的体内化疗增敏作用。在组织芯片中，聚集素在62.5%的肿瘤中表达。停用雌激素和紫杉醇后，MCF-7细胞聚集素表达增加。OGX-011或siRNA Clusterin以剂量依赖、序列特异性的方式将Clusterin水平降低90%，并显著增强体外对紫杉醇的化疗敏感性。当OGX-011或siRNA Clusterin联合使用时，与单独使用紫杉醇相比，IC50降低了2个对数。体内应用OGX-011可增强紫杉醇显著延缓MCF-7肿瘤生长的作用。这些数据确定Clusterin是一个有效的治疗靶点，并为在乳腺癌的多种疗法中测试OGX-011提供了临床前的原则证明。
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 IC5o 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.