Partin A W, Briggman J V, Subong E N, Szaro R, Oreper A, Wiesbrock S, Meyer J, Coffey D S, Epstein J I
Department of Urology, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA.
Urology. 1997 Nov;50(5):800-8. doi: 10.1016/S0090-4295(97)00337-3.
A nuclear matrix protein (PC-1) was previously identified and reported to be present only in human prostate cancer but absent in tissue from the same prostate containing either benign prostatic hyperplasia (BPH) or normal prostate tissue. The PC-1 protein was identified by high resolution two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and exhibited a molecular mass of 56 kDa and an isoelectric point of 6.58. This work investigates the immunohistochemical characterization of PRO:4-216, a monoclonal antibody to PC-1.
Areas of the 2D-PAGE gels containing the human prostate cancer nuclear matrix proteins near PC-1 were isolated, eluted, and injected into mice to develop monoclonal antibodies. Antibodies were screened by immunofluorescence for nuclear reactivity to a human prostate cancer cell line (LnCaP) and by 1D and 2D Western blots for reactivity with prostate cancer nuclear matrix proteins. Monoclonal antibodies from the selected clones were affinity purified. The monoclonal antibody PRO:4-216 was used to analyze frozen tissue from 20 cancerous, 22 BPH, and 22 normal regions from fresh human prostate specimens. Tissue sections were analyzed for their immunohistochemical (IHC) (horseradish peroxidase) staining.
Using a reference value for positive staining at an IHC score of greater than 50, 85% (17 of 20) of the cancerous, 5% (1 of 22) of the BPH, and 9% (2 of 22) of the normal prostate tissues stained positive. The one BPH and two normal tissues that stained positive were taken from prostates in which the adjacent cancerous tissue also demonstrated high IHC scores (greater than 225).
These data demonstrate nuclear reactivity on fresh frozen human prostate cancer tissue for the monoclonal antibody PRO:4-216. PRO:4-216 may aid in distinguishing normal prostate and BPH from cancerous tissue.
先前已鉴定出一种核基质蛋白（PC-1），据报道其仅存在于人类前列腺癌中，而在含有良性前列腺增生（BPH）或正常前列腺组织的同一前列腺组织中不存在。通过高分辨率二维聚丙烯酰胺凝胶电泳（2D-PAGE）鉴定出PC-1蛋白，其分子量为56 kDa，等电点为6.58。本研究调查了针对PC-1的单克隆抗体PRO:4-216的免疫组织化学特征。
分离、洗脱并注射二维聚丙烯酰胺凝胶中靠近PC-1的含有人前列腺癌核基质蛋白的区域，以制备单克隆抗体。通过免疫荧光筛选抗体对人前列腺癌细胞系（LnCaP）的核反应性，并通过一维和二维蛋白质印迹筛选其与前列腺癌核基质蛋白的反应性。对所选克隆的单克隆抗体进行亲和纯化。使用单克隆抗体PRO:4-216分析来自新鲜人类前列腺标本的20个癌组织、22个BPH组织和22个正常组织的冷冻组织。对组织切片进行免疫组织化学（IHC）（辣根过氧化物酶）染色分析。
使用免疫组织化学评分大于50作为阳性染色的参考值，85%（20个中的17个）的癌组织、5%（22个中的1个）的BPH组织和9%（22个中的2个）的正常前列腺组织染色呈阳性。染色呈阳性的1个BPH组织和2个正常组织取自相邻癌组织免疫组织化学评分也较高（大于225）的前列腺。
这些数据表明单克隆抗体PRO:4-216对新鲜冷冻的人类前列腺癌组织具有核反应性。PRO:4-216可能有助于将正常前列腺组织和BPH组织与癌组织区分开来。
