PURIFICATION AND ANALYSIS OF GROWTH-REGULATING PROTEINS SECRETED BY A HUMAN-MELANOMA CELL-LINE

被引:49
作者
APFEL, R [1 ]
LOTTSPEICH, F [1 ]
HOPPE, J [1 ]
BEHL, C [1 ]
DURR, G [1 ]
BOGDAHN, U [1 ]
机构
[1] DEPT NEUROL,TUMORBIOL LAB,JOSEF SCHNEIDER STR 11,W-8700 WURZBURG,GERMANY
关键词
ENDOZEPINE; MALIGNANT MELANOMA; MELANOMA INHIBITING ACTIVITY; BETA-2-MICROGLOBULIN; TIMP-2; UBIQUITIN;
D O I
10.1097/00008390-199212000-00006
中图分类号
R73 [肿瘤学];
学科分类号
100214 ;
摘要
Supernatants of a human malignant cell line established from a CNS metastasis, contained several proteins with putative growth regulating functions. BioGel P-10 gel filtration chromatography, reverse phase HPLC purification, and amino-terminal sequencing of purified peptides resulted in characterization of beta2-microglobulin (beta2M, 10 kD), ubiquitin (6 kD), and tissue inhibitor of metalloproteinases 2 (TIMP-2, 21 kD). In addition, CNBr cleavage and purification of resulting peptides revealed diazepam binding inhibitor (DBI, 8 kD) and melanoma inhibiting activity (MIA, 11 kD). The secretion of beta2M as part of the HLA-class I complex may be related to impaired autologous anti-tumour immune function; ubiquitin may play a role in activation or deactivation of extracellular proteins or cell-cell interactions. As HTZ-19 cells respond in a dose-dependent manner to midozolam, DBI may interfere with growth regulation mediated by diazepam receptor sites. In a collagenolytic assay, TIMP-2 interfered with metalloproteinase functions, which are required for degradation of collagen type IV and organotopic metastasis. MIA is clearly associated with a proliferation inhibiting effect on HTZ-19 cells. In conclusion, although this tumour shows a degree of progression, several proteins with putative functions at different cellular levels were identified, related to proliferation as well as to the type of metastasis.
引用
收藏
页码:327 / 336
页数:10
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