Tissue-based Quantification of 8-Hydroxy-2′-Deoxyguanosine in Human Prostate Biopsies Using Quantitative Fluorescence Imaging Analysis
Tissue-based Quantification of 8-Hydroxy-2′-Deoxyguanosine in Human Prostate Biopsies Using Quantitative Fluorescence Imaging Analysis
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DOI：
10.1016/j.urology.2009.01.052
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发表时间：
2009-11-01
期刊：
影响因子：
2.1
通讯作者：
Hemstreet, George P.
中科院分区：
文献类型：
--
作者：
Richardson, Tim;McCanse, Webb;Hemstreet, George P.
目的：采用一种新的定量荧光成像分析方法，对20例年龄匹配的良性（对照）和含癌组织切片的前列腺活检标本进行8-羟基-2 ′-脱氧鸟苷（8-OHdG）定量。8-在单个腺泡核和周围的基质核内定量OHdG。石蜡切片用RNA酶和蛋白酶处理以暴露核染色质，与结合至饱和的抗8-OHdG小鼠单克隆抗体反应，并用Alex Fluor 488标记的二级山羊抗小鼠IgG检测，并用校准的定量荧光成像分析系统定量。结果进行了分析，使用配对Student的t test.Results 8-OHdG成功地量化内的个别细胞隔室，而不需要激光组织解剖，使用荧光标记的8-OHdG的平均像素强度。8-OHdG的整体表达的配对分析，以及腺泡和间质单独，显示癌和对照前列腺组织之间没有差异。所有前列腺癌患者和良性前列腺病变患者的腺泡内8-OHdG均显著高于周围吻合口（P <0.05）。8-与年龄匹配的良性前列腺组织相比，含癌前列腺组织的腺泡或间质中OHdG没有显著升高。虽然8-OHdG显着升高腺泡核与周围的基质核在含癌和良性前列腺组织，它本身，不是一个强大的生物标志物前列腺癌的风险评估。泌尿学74：1174-1179，2009。(C)2009年由Elsevier Inc.出版
OBJECTIVES To quantify 8-hydroxy-2'-deoxyguanosine (8-OHdG) in prostate stromal and acini tissue compartments from benign and cancer-containing prostate specimens using a new quantitative fluorescence imaging analysis protocol.METHODS Prostate biopsy specimens from 20 age-matched benign (control) and cancer-containing tissue sections were used to quantify 8-OHdG. 8-OHdG was quantified within individual acini nuclei and the surrounding stroma nuclei. Paraffin sections were treated with RNAse and protease to expose the nuclear chromatin, reacted with anti-8-OHdG mouse monoclonal antibody bound to saturation and detected with secondary goat anti-mouse IgG labeled with Alex Fluor 488, and quantified with a calibrated quantitative fluorescence imaging analysis system. The results were analyzed using a paired Student's t test.RESULTS 8-OHdG was successfully quantified within individual cellular compartments without the need for laser tissue dissection, using the mean pixel intensity of fluorescent-labeled 8-OHdG. Matched-pair analysis of the global expression of 8-OHdG, as well as the acini and stroma individually, revealed no difference between the cancerous and control prostatic tissue. All patients with prostate cancer and those with benign findings had significantly greater 8-OHdG within the acini compared with the surrounding stoma (P < .05).CONCLUSIONS A protocol to quantify 8-OHdG in paraffin-embedded human prostatic tissue was successfully developed. 8-OHdG was not significantly elevated in the acini or stroma of cancer-containing prostatic tissue compared, with age-matched benign prostatic tissue. Although 8-OHdG was significantly elevated in the acini nuclei compared with the surrounding stroma nuclei in both cancer-containing and benign prostatic tissue, it, by itself, was not a strong biomarker for prostate cancer risk assessment. UROLOGY 74: 1174-1179, 2009. (C) 2009 Published by Elsevier Inc.