Department of Oncology, Weatherall Institute of Molecular Medicine, University of Oxford, John Radcliffe Hospital, Oxford, UK.
Mol Ther Nucleic Acids. 2012 Jun 19;1(6):e30. doi: 10.1038/mtna.2012.19.
RNA-directed antisense and interference therapeutics are a promising treatment option for cancer. The demonstration of depletion of target proteins within human tumors in vivo using validated methodology will be a key to the application of this technology. Here, we present a flow cytometric-based approach to quantitatively determine protein levels in solid tumor material derived by fiber optic brushing (FOB) of non-small cell lung cancer (NSCLC) patients. Focusing upon the survivin protein, and its depletion by an antisense oligonucleotide (ASO) (LY2181308), we show that we can robustly identify a subpopulation of survivin positive tumor cells in FOB samples, and, moreover, detect survivin depletion in tumor samples from a patient treated with LY2181308. Survivin depletion appears to be a result of treatment with this ASO, because a tumor treated with conventional cytotoxic chemotherapy did not exhibit a decreased percentage of survivin positive cells. Our approach is likely to be broadly applicable to, and useful for, the quantification of protein levels in tumor samples obtained as part of clinical trials and studies, facilitating the proof-of-principle testing of novel targeted therapies.
RNA 导向的反义与干扰疗法是癌症治疗的一种很有前途的选择。使用经验证的方法在体内证明目标蛋白在人类肿瘤中的耗竭将是这项技术应用的关键。在这里，我们提出了一种基于流式细胞术的方法，用于定量测定通过纤维光学刷取（FOB）非小细胞肺癌（NSCLC）患者获得的实体瘤材料中的蛋白质水平。我们专注于生存素蛋白及其反义寡核苷酸（ASO）（LY2181308）的耗竭，结果表明我们可以在 FOB 样本中稳健地鉴定出生存素阳性肿瘤细胞的亚群，并且，还可以检测到用 LY2181308 治疗的患者的肿瘤样本中生存素的耗竭。生存素的耗竭似乎是由于用这种 ASO 治疗的结果，因为用常规细胞毒性化疗治疗的肿瘤没有表现出生存素阳性细胞百分比的降低。我们的方法可能广泛适用于并有助于定量测定临床试验和研究中获得的肿瘤样本中的蛋白质水平，从而促进新型靶向治疗的原理验证测试。
