Diversity of melanoma plasma membrane proteinases: inhibition of collagenolytic and cytolytic activities by minocycline.
Diversity of melanoma plasma membrane proteinases: inhibition of collagenolytic and cytolytic activities by minocycline.
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DOI：
10.1093/jnci/75.3.517
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发表时间：
1985-09
期刊：
影响因子：
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通讯作者：
S. Zucker;R. Lysik;N. Ramamurthy;L. Golub;J. Wieman;D. Wilkie
中科院分区：
文献类型：
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作者：
S. Zucker;R. Lysik;N. Ramamurthy;L. Golub;J. Wieman;D. Wilkie
检查了来自 C57BL/6 小鼠的 B16-BL6 黑色素瘤细胞和亚细胞部分的组织破坏性蛋白酶。在氮空化后使用蔗糖密度梯度离心分离癌细胞细胞器。使用放射性标记的蛋白质和合成底物对丝氨酸、半胱氨酸和金属蛋白酶进行测定。通过测量与黑色素瘤细胞共培养的 59Fe 标记红细胞 (RBC) 释放的同位素，对肿瘤诱导的红细胞裂解进行定量；红细胞来自 Wistar 大鼠。在上述测定中使用对不同类别的蛋白酶具有特异性的酶抑制剂来对负责底物降解的酶进行分类。结果表明，完整的黑色素瘤细胞、细胞器和细胞质含有蛋白酶，可以降解胶原蛋白和明胶并裂解正常红细胞。黑色素瘤质膜富含胶原酶、明胶酶、半胱氨酸蛋白酶、纤溶酶原激活剂和细胞溶解活性。 EDTA 和 1,10-菲咯啉对肿瘤胶原溶解、明胶溶解和细胞溶解活性的抑制作用，但二异丙基氟磷酸盐和 N α-对甲苯磺酰基-L-赖氨酸氯甲基酮没有抑制作用，表明金属蛋白酶是这些测定中的活性酶。米诺环素是一种合成四环素，对其他哺乳动物胶原酶具有明显的抑制活性，也能抑制黑色素瘤的胶原溶解和细胞溶解活性。
The tissue-destructive proteinases of B16-BL6 melanoma cells from C57BL/6 mice and subcellular fractions were examined. Cancer cell organelles were isolated following nitrogen cavitation with the use of sucrose density gradient centrifugation. Serine, cysteine, and metalloproteinases were assayed with the use of radiolabeled proteins and synthetic substrates. Tumor-induced red blood cell lysis was quantitated by measurement of the release of isotope from 59Fe-labeled red blood cells (RBC) cocultivated with melanoma cells; the RBC were from Wistar rats. Enzyme inhibitors with specificity toward different classes of proteinases were used in the above assays to categorize the enzymes responsible for substrate degradation. Results indicated that intact melanoma cells, cell organelles, and cytosol contain proteinases that can degrade collagen and gelatin and lyse normal RBC. Melanoma plasma membranes are highly enriched in collagenase, gelatinase, cysteine proteinase, plasminogen activator, and cytolytic activity. The inhibition of tumor collagenolytic, gelatinolytic, and cytolytic activities by EDTA and 1,10-phenanthroline but not by diisopropyl fluorophosphate and N alpha-p-tosyl-L-lysine chloromethyl ketone indicates that metalloproteinases are the active enzymes in these assays. Minocycline, a synthetic tetracycline with demonstrable inhibitory activity with other mammalian collagenases, also inhibited melanoma collagenolytic and cytolytic activities.