Hernberg M, Turunen J P, von Boguslawsky K, Muhonen T, Pyrhönen S
Department of Internal Medicine, University of Helsinki, Finland.
Melanoma Res. 1998 Jun;8(3):283-91. doi: 10.1097/00008390-199806000-00013.
Biopsy specimens from 12 patients with metastatic melanoma were longitudinally analysed to evaluate changes in proliferation activity and CD4+/CD8+ ratios during the course of the disease. The primary tumours of the patients who subsequently had metastatic disease were also each matched with tumours from two controls whose disease remained localized, and were compared with regard to tumour proliferation. Immunohistochemistry was performed using the avidin-biotin complex (ABC) immunoperoxidase technique, using bcl-2, p53, mdm-2 and Ki-67 as the primary monoclonal antibodies, and the percentage of positively stained melanocytic cells was calculated. Frozen sections were also available from metastatic lesions excised from eight of our patients before treatment initiation and at the time of disease progression. These specimens were prepared for microscopy, and quantitative characterization of CD4+ (OKT 4a) and CD8+ (OKT 8) cells was performed. Compared with the localized melanomas bcl-2 expression was higher in those primary melanomas that later metastasized (P = 0.068, Wilcoxon; P = 0.038, median test). Mdm-2 and Ki-67 expression did not differ in the primary tumours of patients and controls, but a statistically significant trend was observed towards increasing expression with the progression of the disease (two-sided exact P-values: 0.04 and 0.05, respectively). Patients with a low Ki-67 index in their first metastasis had a better prognosis when compared with patients with high indexes (P = 0.008, log-rank). Furthermore, most patients with decreasing CD4+/CD8+ ratios had increasing p53 immunoreactivity. Our findings suggest that Ki-67 and bcl-2 may be useful for predicting the prognosis of melanoma patients. Mdm-2 is a new but promising marker in melanoma and deserves further evaluation.
对12例转移性黑色素瘤患者的活检标本进行纵向分析，以评估疾病过程中增殖活性和CD4⁺/CD8⁺比值的变化。随后发生转移性疾病的患者的原发性肿瘤也分别与两名疾病仍局限的对照患者的肿瘤进行匹配，并就肿瘤增殖情况进行比较。采用抗生物素蛋白-生物素复合物（ABC）免疫过氧化物酶技术进行免疫组织化学检测，以bcl-2、p53、mdm-2和Ki-67作为主要单克隆抗体，并计算阳性染色黑素细胞的百分比。我们的8例患者在开始治疗前和疾病进展时切除的转移性病变也有冰冻切片。这些标本经处理用于显微镜检查，并对CD4⁺（OKT 4a）和CD8⁺（OKT 8）细胞进行定量表征。与局限性黑色素瘤相比，后来发生转移的原发性黑色素瘤中bcl-2表达更高（Wilcoxon检验，P = 0.068；中位数检验，P = 0.038）。患者和对照的原发性肿瘤中mdm-2和Ki-67表达无差异，但随着疾病进展观察到表达有统计学意义的增加趋势（双侧精确P值分别为0.04和0.05）。首次转移时Ki-67指数低的患者与高指数患者相比预后更好（P = 0.008，对数秩检验）。此外，大多数CD4⁺/CD8⁺比值降低的患者p53免疫反应性增加。我们的研究结果表明，Ki-67和bcl-2可能有助于预测黑色素瘤患者的预后。mdm-2是黑色素瘤中的一种新的但有前景的标志物，值得进一步评估。
