COEXPRESSION OF VIMENTIN AND KERATINS BY HUMAN-MELANOMA TUMOR-CELLS - CORRELATION WITH INVASIVE AND METASTATIC POTENTIAL
COEXPRESSION OF VIMENTIN AND KERATINS BY HUMAN-MELANOMA TUMOR-CELLS - CORRELATION WITH INVASIVE AND METASTATIC POTENTIAL
复制标题
DOI：
10.1093/jnci/84.3.165
复制
发表时间：
1992-02-05
影响因子：
10.3
通讯作者：
STETLERSTEVENSON, W
中科院分区：
文献类型：
--
作者：
HENDRIX, MJC;SEFTOR, EA;STETLERSTEVENSON, W
背景：包括静脉溶酶蛋白在内的几种蛋白质标记物已被用于诊断人类黑色素瘤。因为黑色素瘤通常在诊断时已经转移，因此需要确定转移潜力的早期预后标记。通常，波形蛋白存在于间充质细胞中，角蛋白存在于上皮细胞中，但最近的研究报道了在上皮和非上皮肿瘤（包括一些黑色素瘤）中波形蛋白和角蛋白的共表达。目的：我们的目的是确定波形蛋白和角蛋白的共表达是否与肿瘤细胞的侵袭和转移行为相关。方法：我们评估了9种表达vimentin和其他侵袭性肿瘤行为标志物（HMB-45、S-100、HLA-ABC类和HLA-DR类组织相容性抗原，以及K8和K18角蛋白）的人黑色素瘤细胞系。在高度转移的C8161细胞系、低转移的A375P细胞系和中度转移的A375M细胞系中检测K8和K18角蛋白的水平。为了确定角蛋白的存在是否会影响迁移能力，我们通过转染突变的K18互补DNA改变了C8161细胞角蛋白丝的构象结构。我们还测定了人类IV型胶原酶的信使RNA水平，这是一种侵袭和转移的酶标记物。结果：在A375P细胞中，考马斯氏染色凝胶双向电泳、免疫印迹和免疫荧光染色均未检测到K8或K18的水平。Western和Northern印迹法检测A375M细胞显示低水平的K8和K18，具有独特的荧光亚群。相比之下，C8161细胞的K8和K18水平在所有细胞中都很高。IV型胶原酶信使RNA水平在A375P细胞中最低，在C8161细胞中最高，这与体外侵袭能力和胸腺裸鼠的转移潜力有关。来自C8161亲本系的C1070-10和C1070-14表现出明显的形态变化，角蛋白丝断裂，侵袭和转移潜力降低，与迁移活性降低直接相关。结论：这些发现表明vimentin与K8和K18角蛋白的共表达与三种具有代表性的人类黑色素瘤细胞系的侵袭和转移行为有关。
Background: Several protein markers, including vimentin, have been used to diagnose human melanoma. Because melanoma often has metastasized by the time of diagnosis, early markers prognostic for metastatic potential need to be identified. Commonly, vimentin is found in mesenchymal cells, and keratins are present in epithelial cells, but recent studies report coexpression of vimentin and keratin(s) in epithelial and nonepithelial neoplasms, including some melanomas. Purpose: Our purpose was to determine whether coexpression of vimentin and keratin(s) is correlated with tumor cell invasion and metastatic behavior. Methods: We evaluated nine human melanoma cell lines expressing vimentin and other markers of aggressive tumor behavior (HMB-45, S-100, HLA-ABC class I and HLA-DR class II histocompatibility antigens, and K8 and K18 keratins). Levels of K8 and K18 keratins were determined in the highly metastatic C8161 cell line, the poorly metastatic A375P line, and the moderately metastatic A375M line. To determine whether the presence of keratin affects migratory ability, we altered the conformational structure of keratin filaments in C8161 cells by transfection with a mutant K18 complementary DNA. We also determined messenger RNA levels of human type IV collagenase, an enzyme marker for invasion and metastasis. Results: In A375P cells, two-dimensional electrophoresis with Coomassie-stained gels, immunoblotting, and immunofluorescence staining showed no detectable levels of K8 or K18. A375M cells showed low levels of K8 and K18 by Western and Northern blotting, with a distinctive fluorescent subpopulation of cells. In comparison, K8 and K18 levels in C8161 cells were high in all cells. Type IV collagenase messenger RNA levels were lowest in A375P cells and highest in C8161 cells, correlating with invasive ability in vitro and metastatic potential in athymic nude mice. The transfectant clones C1070-10 and C1070-14 derived from the C8161 parent line showed dramatic morphological changes, disrupted keratin filaments, and decreased invasive and metastatic potential directly correlated with a reduction in migratory activity. Conclusion: These findings show a correlation between the coexpression of vimentin with K8 and K18 keratins and the invasive and metastatic behavior of three representative human melanoma cell lines.