Tsuji Tsutomu, Kawada Yoko, Kai-Murozono Mieko, Komatsu Shinya, Han Seon Ae, Takeuchi Ken-ichi, Mizushima Hiroto, Miyazaki Kaoru, Irimura Tatsuro
Department of Microbiology, Hoshi University, Tokyo, Japan.
Clin Exp Metastasis. 2002;19(2):127-34. doi: 10.1023/a:1014573204062.
We evaluated the role of soluble factors produced from epidermal cells in melanoma cell motility by using the Boyden chamber chemoinvasion system. The migration of two melanoma cell lines, A375 and Mewo, was potentiated by conditioned media of A431 epidermoid cells in a concentration-dependent manner. The enhancement of A375 melanoma cell motility induced by the conditioned medium was blocked by antibodies against either alpha3 or beta1 integrin subunit. The motility-stimulating activity was recovered in the same fraction as the alpha3 integrin-dependent adhesion-promoting activity in a high-molecular-weight (>200 kDa) fraction on Superose 12 gel chromatography, and adsorbed with an anti-laminin-5 antibody. Purified laminin-5 was capable of potentiating melanoma cell migration as measured in either the chemotaxis assay with a soluble form of laminin-5 or the haptotaxis assay with membranes coated with a mixture of laminin-5 and Matrigel. Furthermore, immobilized laminin-5 induced A375 melanoma cells to secrete matrix metalloproteinase-9 (type IV collagenase) into the culture medium. These results strongly suggest that the interaction of laminin-5 produced in the epidermis with alpha3beta1 integrin on melanoma cells is involved in cell migration, invasion, and degradation of extracellular matrix proteins.
我们通过使用博伊登室化学侵袭系统评估了表皮细胞产生的可溶性因子在黑色素瘤细胞运动中的作用。两种黑色素瘤细胞系A375和Mewo的迁移受到A431表皮样细胞条件培养基的浓度依赖性增强。条件培养基诱导的A375黑色素瘤细胞运动增强被针对α3或β1整合素亚基的抗体所阻断。在Superose 12凝胶过滤色谱中，运动刺激活性与高分子量（>200 kDa）部分中α3整合素依赖性黏附促进活性存在于同一组分中，并可被抗层粘连蛋白-5抗体吸附。纯化的层粘连蛋白-5能够增强黑色素瘤细胞迁移，这在使用可溶性层粘连蛋白-5的趋化性测定或使用涂有层粘连蛋白-5和基质胶混合物的膜的触觉趋化性测定中均有体现。此外，固定化的层粘连蛋白-5诱导A375黑色素瘤细胞向培养基中分泌基质金属蛋白酶-9（IV型胶原酶）。这些结果强烈表明，表皮中产生的层粘连蛋白-5与黑色素瘤细胞上的α3β1整合素之间的相互作用参与了细胞迁移、侵袭以及细胞外基质蛋白的降解。
