Richardson Tim, McCanse Webb, Casale George P, Huang Dali, Tian Jennifer, Elkahwaji Johny E, Lele Subodh, Hemstreet George P
Department of Urology, University of Nebraska Medical Center and Omaha Veterans Affairs Hospital, Omaha, Nebraska 68114, USA.
Urology. 2009 Nov;74(5):1174-9. doi: 10.1016/j.urology.2009.01.052. Epub 2009 Aug 29.
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.
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.
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).
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.
使用一种新的定量荧光成像分析方案，对来自良性和含癌前列腺标本的前列腺基质和腺泡组织区室中的8-羟基-2'-脱氧鸟苷（8-OHdG）进行定量。
使用来自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对8-OHdG进行定量。在单个腺泡细胞核和周围的基质细胞核内对8-OHdG进行定量。石蜡切片用核糖核酸酶和蛋白酶处理以暴露核染色质，与饱和结合的抗8-OHdG小鼠单克隆抗体反应，并用Alex Fluor 488标记的山羊抗小鼠IgG二抗进行检测，并用校准的定量荧光成像分析系统进行定量。结果采用配对学生t检验进行分析。
使用荧光标记的8-OHdG的平均像素强度，无需激光组织切割即可在单个细胞区室内成功定量8-OHdG。对8-OHdG的整体表达以及腺泡和基质分别进行配对分析，结果显示癌性前列腺组织和对照前列腺组织之间无差异。所有前列腺癌患者和良性病变患者的腺泡内8-OHdG均显著高于周围的基质（P < 0.05）。
成功开发了一种对石蜡包埋的人前列腺组织中8-OHdG进行定量的方案。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基质中8-OHdG没有显著升高。尽管在含癌和良性前列腺组织中，腺泡细胞核内的8-OHdG均显著高于周围的基质细胞核，但它本身并不是前列腺癌风险评估的强生物标志物。
