Survivin antisense oligonucleotides effectively radiosensitize colorectal cancer cells in both tissue culture and murine xenograft models
Survivin antisense oligonucleotides effectively radiosensitize colorectal cancer cells in both tissue culture and murine xenograft models
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DOI：
10.1016/j.ijrobp.2008.02.011
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发表时间：
2008-05-01
影响因子：
7
通讯作者：
Roedel, Claus
中科院分区：
文献类型：
--
作者：
Roedel, Franz;Frey, Benjamin;Roedel, Claus
目的：Survivin在结直肠癌中显示放射抵抗因子。在本研究中，我们确定直肠癌患者的survivin信使RNA水平是否能预测新辅助放化疗后的肿瘤反应，以及使用反义寡核苷酸（ASOs）抑制survivin是否能增强放射反应。方法与材料：用survivin ASO （LY2181308）转染SW480结直肠癌细胞，以0 ~ 8 Gy剂量照射。通过免疫印迹、细胞计数和caspase 3/7活性检测Survivin表达、细胞周期分布、γ - H2AX荧光和凋亡诱导。克隆成活通过集落形成试验测定。采用SW480异种移植瘤模型，研究survivin衰减和照射对肿瘤生长的影响。此外，通过Affymetrix微阵列分析，研究了患者活检标本中的survivin信使RNA水平。结果：在20例直肠癌患者的转化研究中，survivin水平升高与肿瘤局部复发风险显著升高相关（p = 0.009）。用survivin ASOs和照射处理SW480细胞，导致凋亡细胞百分比、caspase 3/7活性、G(2)/M期细胞比例和H2AX磷酸化增加。与对照处理的细胞相比，克隆存活率降低。此外，SW480异种移植物的治疗；ASOs和放疗显著延缓了肿瘤的生长。结论：Survivin可能是直肠癌患者的一种分子生物标志物。此外，体外和体内数据表明，survivin作为一种分子靶点具有潜在的作用，可以改善直肠癌患者对放疗的治疗反应。(c) 2008爱思唯尔公司
Purpose: Survivin shows a radiation resistance factor in colorectal cancer. In the present study, we determined whether survivin messenger RNA levels in patients with rectal cancer predict tumor response after neoadjuvant radiochemotherapy and whether inhibition of survivin by the use of antisense oligonucleotides (ASOs) enhances radiation responses.Methods and Materials: SW480 colorectal carcinoma cells were transfected with survivin ASO (LY2181308) and irradiated with doses ranging from 0-8 Gy. Survivin expression, cell-cycle distribution, gamma H2AX fluorescence, and induction of apoptosis were monitored by means of immunoblotting, How cytometry, and caspase 3/7 activity. Clonogenic survival was determined by using a colony-forming assay. An SW480 xenograft model was used to investigate the effect of survivin attenuation and irradiation on tumor growth. Furthermore, survivin messenger RNA levels were studied in patient biopsy specimens by using Affymetrix microarray analysis.Results: In the translational study of 20 patients with rectal cancer, increased survivin levels were associated with significantly greater risk of local tumor recurrence (p = 0.009). Treatment of SW480 cells with survivin ASOs and irradiation resulted in an increased perceentage of apoptotic cells, caspase 3/7 activity, fraction of cells in the G(2)/M phase, and H2AX phosphorylation. Clonogenic survival decreased compared with control-treated cells. Furthermore, treatment of SW480 xenografts; with survivin ASOs and irradiation resulted in a significant delay in tumor growth.Conclusion: Survivin appears to be a molecular biomarker in patients with rectal cancer. Furthermore, in vitro and in vivo data suggest a potential role of survivin as a molecular target to improve treatment response to radiotherapy in patients with rectal cancer. (c) 2008 Elsevier Inc.