LOCALIZATION OF CATHEPSIN-B IN 2 HUMAN LUNG-CANCER CELL-LINES

被引:61
作者
ERDEL, M
TREFZ, G
SPIESS, E
HABERMAAS, S
SPRING, H
LAH, T
EBERT, W
机构
[1] GERMAN CANC RES CTR, INST CELL & TUMOR BIOL, NEUENHEIMER FELD 280, W-6900 HEIDELBERG 1, GERMANY
[2] THORAX KLIN HEIDELBERG ROHRBACH, W-6900 HEIDELBERG, GERMANY
[3] GERMAN CANC RES CTR, INST EXPTL PATHOL, W-6900 HEIDELBERG 1, GERMANY
[4] J STEFAN INST, YU-6111 LJUBLJANA, YUGOSLAVIA
关键词
Cathepsin B; immunocytochemistry; laser scanning microscopy; localization; plasma membrane; tumor cells;
D O I
10.1177/38.9.2201737
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
We demonstrated the cysteine proteinase cathepsin B in two human lung tumor cell lines by cytochemical and immunocytochemical methods. The cell lines were derived from a squamous cell carcinoma of the lung (HS-24) and a metastasis to the adrenal gland from an adenocarcinoma of the lung (SB-3). For comparison and control, normal human lung fibroblast cells (Wi-38) were also investigated. Intracellular cathepsin B activity was detected in all three cell lines. SB-3 and the normal fibroblast cells showed almost equal cathepsin B activity, which was considerably stronger than that in the HS-24 cells. Specific inhibitors for cathepsin B (E64, leupeptin, antipain) suppressed its activity completely. Stefin A, the physiological cathepsin B inhibitor, was less effective; this might depend on its limited penetrability into living cells. Localization of the cathepsin B was performed by conventional immunofluorescence microscopy and laser scanning microscopy. With specific anti-cathepsin B antibodies, the enzyme was localized in HS-24, SB-3, and Wi-38 fibroblast cells within perinuclear granules representing the lysosomal compartment. In the SB-3 cells, we additionally localized a minor fraction of the enzyme bound to the plasma membrane in a speckled distribution, accessible to the antibodies from the outside. This direct demonstration of cathepsin B distribution supports biochemical data about the dual localization of the enzyme in tumor cells. It also supports the possibility of a direct involvement of cathepsin B in the degradation of the extracellular matrix, and thus a contribution of the enzyme in invasion and metastasis.
引用
收藏
页码:1313 / 1321
页数:9
相关论文
共 39 条
[1]  
Barrett A. J., 1977, PROTEINASES MAMMALIA, P181
[2]  
BARRETT AJ, 1981, METHOD ENZYMOL, V80, P535
[3]   CATHEPSIN-B1 - LYSOSOMAL ENZYME THAT DEGRADES NATIVE COLLAGEN [J].
BURLEIGH, MC ;
BARRETT, AJ ;
LAZARUS, GS .
BIOCHEMICAL JOURNAL, 1974, 137 (02) :387-+
[4]   FLUORESCENT ASSAY OF PROTEINASES IN CULTURED MAMMALIAN-CELLS [J].
DOLBEARE, F ;
VANDERLAAN, M .
JOURNAL OF HISTOCHEMISTRY & CYTOCHEMISTRY, 1979, 27 (11) :1493-1495
[5]  
DOLBEARE FA, 1977, CLIN CHEM, V23, P1485
[6]   FURTHER-STUDIES ON ACTIVATION OF PROCOLLAGENASE, LATENT PRECURSOR OF BONE COLLAGENASE - EFFECTS OF LYSOSOMAL CATHEPSIN-B, PLASMIN AND KALLIKREIN, AND SPONTANEOUS ACTIVATION [J].
EECKHOUT, Y ;
VAES, G .
BIOCHEMICAL JOURNAL, 1977, 166 (01) :21-31
[7]   USE OF THE AVIDIN-BIOTIN-PEROXIDASE COMPLEX (ABC) METHOD FOR THE LOCALIZATION OF RABBIT CATHEPSIN B IN CELLS AND TISSUES [J].
GRAF, FM ;
STRAULI, P .
JOURNAL OF HISTOCHEMISTRY & CYTOCHEMISTRY, 1983, 31 (06) :803-810
[8]  
HANEWINKEL H, 1987, J BIOL CHEM, V262, P12351
[9]   THE DISTRIBUTION OF CATHEPSIN-B IN HUMAN-TISSUES [J].
HOWIE, AJ ;
BURNETT, D ;
CROCKER, J .
JOURNAL OF PATHOLOGY, 1985, 145 (04) :307-314
[10]  
KEREN Z, 1988, CANCER RES, V48, P1416