Kobayashi T, Hayashi A, Ura-Ishikou A, Tajima S, Nishikawa T
Department of Dermatology and Clinical Investigation, Tokyo Daini National Hospital, Japan.
Cancer Lett. 1994 Oct 14;85(2):165-9. doi: 10.1016/0304-3835(94)90270-4.
Mouse B16 melanoma cells are of heterogenous populations and consist of melanotic and amelanotic cells. Two clones, M3 (melanotic) and A7 (amelanotic), were screened from B16 melanoma cells using a microscope, and gelatinolytic activities in these clones were studied with zymography. Gelatinolytic enzyme was found in the cultured medium of A7 cells, but not in M3 cells. The enzymes with apparent molecular weights of 70, 140 and 280 kDa were detected and found to form complexes: a dimer or more than dimer. Their enzyme activities were not inhibited by ethylenediaminetetraacetic acid. The optimal pH of this enzyme was found to be at neutral pH. Preferential expression of this novel gelatinolytic enzyme in amelanotic melanoma will be related to tumorigenic and metastatic phenotype of amelanotic melanoma.
小鼠B16黑色素瘤细胞是异质群体，由黑色素细胞和无黑色素细胞组成。使用显微镜从B16黑色素瘤细胞中筛选出两个克隆，M3（黑色素细胞）和A7（无黑色素细胞），并通过酶谱法研究这些克隆中的明胶酶解活性。在A7细胞的培养基中发现了明胶酶解酶，但在M3细胞中未发现。检测到表观分子量为70、140和280 kDa的酶，并发现它们形成复合物：二聚体或多于二聚体。它们的酶活性不受乙二胺四乙酸抑制。发现这种酶的最佳pH值为中性pH。这种新型明胶酶解酶在无黑色素黑色素瘤中的优先表达将与无黑色素黑色素瘤的致瘤和转移表型相关。
