Li G, Passebosc-Faure K, Lambert C, Gentil-Perret A, Blanc F, Oosterwijk E, Mosnier J F, Genin C, Tostain J
Department of Urology, North Hospital, CHU of Saint-Etienne, France.
Clin Cancer Res. 2001 Jan;7(1):89-92.
Monoclonal antibody (mAb) G250 is a well characterized and specific mAb to renal cell carcinoma (RCC). The gene G250 was recently cloned and was proved to be homologous to MN/CA9. The G250/MN/CA9 antigen was recently explored as a potential marker for RCC. Flow cytometry (FCM) allows quantitative analysis of cells. The present study describes a flow cytometric method to detect this antigen in human cell lines and in malignant and normal renal tissues. Twelve human carcinoma cell lines (HeLa, Colo205, HT29, BxPC3, OVCAR3, SKOV3, ACHN, A704, CAKI-2, SKRC-59, SKRC-10, and SKRC-52), 10 specimens of normal peripheral blood mononuclear cells, and 38 malignant and 36 adjacent normal renal tissues were studied. The malignant and normal renal tissues were disaggregated mechanically into a single-cell suspension, stained by mAb G250, and analyzed by FCM. All 22 of the clear cell carcinomas, 6 of 8 mixed cell carcinomas, and 3 of 6 granular cell carcinomas were positive for G250/MN/CA9 antigen. SKRC-52 and SKRC-10 were strongly positive for G250/ MN/CA9. The G250/MN/CA9 antigen could also be detected in HeLa, SKOV3, HT29, and A704 cells. One chromophobic, one chromophilic cell carcinoma, the normal renal tissues, and normal peripheral blood mononuclear cells were considered as negative. Our results further confirmed that the G250/MN/CA9 antigen was an ideal marker for RCC, especially for clear cell carcinomas, and that this antigen was present in several types of malignant cells. FCM may serve as a fast tool of immunocytochemical detection of renal cancer cells. Flow cytometric detection of renal cancer cells by using mAb G250 should be further explored.
单克隆抗体（mAb）G250是一种针对肾细胞癌（RCC）的特性明确且特异的单克隆抗体。基因G250最近被克隆出来，并且被证明与MN/CA9同源。G250/MN/CA9抗原最近被探索作为肾细胞癌的一种潜在标志物。流式细胞术（FCM）可对细胞进行定量分析。本研究描述了一种通过流式细胞术检测人细胞系以及恶性和正常肾组织中该抗原的方法。研究了12种人癌细胞系（HeLa、Colo205、HT29、BxPC3、OVCAR3、SKOV3、ACHN、A704、CAKI - 2、SKRC - 59、SKRC - 10和SKRC - 52）、10份正常外周血单个核细胞样本以及38份恶性肾组织和36份相邻正常肾组织。将恶性和正常肾组织机械解离成单细胞悬液，用单克隆抗体G250染色，然后通过流式细胞术进行分析。所有22例透明细胞癌、8例混合细胞癌中的6例以及6例颗粒细胞癌中的3例G250/MN/CA9抗原呈阳性。SKRC - 52和SKRC - 10对G250/MN/CA9呈强阳性。在HeLa、SKOV3、HT29和A704细胞中也能检测到G250/MN/CA9抗原。1例嫌色细胞癌、1例嗜色细胞癌、正常肾组织以及正常外周血单个核细胞被视为阴性。我们的结果进一步证实，G250/MN/CA9抗原是肾细胞癌尤其是透明细胞癌理想的标志物，并且该抗原存在于几种类型的恶性细胞中。流式细胞术可作为检测肾癌细胞的一种快速免疫细胞化学工具。利用单克隆抗体G250进行肾癌细胞的流式细胞术检测值得进一步探索。
