Bakalian Silvin, Marshall Jean Claude, Faingold Dana, Logan Patrick, Antecka Emilia, Burnier Miguel N
McGill University/Henry C. Witelson Ocular Pathology Laboratory, McGill University, Montreal, Quebec, Canada.
Melanoma Res. 2007 Oct;17(5):284-90. doi: 10.1097/CMR.0b013e3282eeea5a.
Uveal melanoma (UM) is the most common malignant intraocular tumor in adults. Despite the high accuracy of clinical diagnosis and advances in local treatment, more than 50% of UM patients develop metastasis within 10 years of initial diagnosis. NM23 is one of the human metastasis suppressor genes. Reduced nm23-H1 expression is correlated with high metastatic potential in many different cancers. The purpose of this study is to investigate the expression of nm23-H1 in UM and its potential value as a prognostic marker. Immunostaining of nm23-H1 was verified in five human UM cell lines with different metastatic potentials. The expression level of nm23-H1 mRNA was evaluated with one-step quantitative real-time PCR. The invasion ability of the cell lines was assessed before and after silencing nm23-H1 with small interference RNA. Thirty-two cases of paraffin-embedded specimens of human UM were immunostained with nm23-H1 monoclonal antibody. The immunostaining was evaluated in a semiquantitative fashion based on extent and intensity. The real-time PCR results of five human UM cell lines showed that expression of nm23-H1 was higher in cell lines with low metastatic potential compared with those with high metastatic potential (P<0.05). The invasive ability of the UM cell lines increased after silencing nm23-H1 expression with small interference RNA (P<0.05). The immunostaining of nm23-H1 was cytoplasmic in all cell lines and UM patients samples. The increased immunostaining intensity of nm23-H1 in patients' samples was associated with better survival rate (Kaplan-Meier test P=0.0097). The expression of nm23-H1 was not correlated with other prognostic factors. It can be concluded that nm23-H1 may be a prognostic marker to predict the survival rate of UM patients and it has the potential to identify high-risk patients.
葡萄膜黑色素瘤(UM)是成人中最常见的眼内恶性肿瘤。尽管临床诊断准确性高且局部治疗取得了进展,但超过50%的UM患者在初次诊断后10年内会发生转移。NM23是人类转移抑制基因之一。nm23-H1表达降低与许多不同癌症的高转移潜能相关。本研究的目的是调查nm23-H1在UM中的表达及其作为预后标志物的潜在价值。在五种具有不同转移潜能的人UM细胞系中验证了nm23-H1的免疫染色。用一步定量实时PCR评估nm23-H1 mRNA的表达水平。在用小干扰RNA沉默nm23-H1前后评估细胞系的侵袭能力。用nm23-H1单克隆抗体对32例人UM石蜡包埋标本进行免疫染色。基于范围和强度以半定量方式评估免疫染色。五个人UM细胞系的实时PCR结果显示,与高转移潜能的细胞系相比,低转移潜能的细胞系中nm23-H1的表达更高(P<0.05)。用小干扰RNA沉默nm23-H1表达后,UM细胞系的侵袭能力增加(P<0.05)。在所有细胞系和UM患者样本中,nm23-H1的免疫染色均为细胞质。患者样本中nm23-H1免疫染色强度增加与更好的生存率相关(Kaplan-Meier检验P=0.0097)。nm23-H1的表达与其他预后因素无关。可以得出结论,nm23-H1可能是预测UM患者生存率的预后标志物,并且有潜力识别高危患者。