LOCALIZATION OF CATHEPSIN-B IN 2 HUMAN LUNG-CANCER CELL-LINES
LOCALIZATION OF CATHEPSIN-B IN 2 HUMAN LUNG-CANCER CELL-LINES
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DOI：
10.1177/38.9.2201737
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发表时间：
1990-09-01
影响因子：
3.2
通讯作者：
EBERT, W
中科院分区：
文献类型：
--
作者：
ERDEL, M;TREFZ, G;EBERT, W
我们通过细胞化学和免疫细胞化学方法在两种人肺肿瘤细胞系中证明了半胱氨酸蛋白酶组织蛋白酶 B。这些细胞系源自肺鳞状细胞癌 (HS-24) 和肺腺癌 (SB-3) 的肾上腺转移瘤。为了进行比较和对照，还研究了正常人肺成纤维细胞（Wi-38）。在所有三种细胞系中均检测到细胞内组织蛋白酶 B 活性。 SB-3和正常成纤维细胞表现出几乎相同的组织蛋白酶B活性，比HS-24细胞中的活性强得多。组织蛋白酶 B 的特异性抑制剂（E64、亮肽素、抗痛剂）完全抑制其活性。 Stefin A（生理组织蛋白酶 B 抑制剂）效果较差；这可能取决于其对活细胞的有限渗透性。通过传统的免疫荧光显微镜和激光扫描显微镜进行组织蛋白酶 B 的定位。使用特异性抗组织蛋白酶 B 抗体，该酶定位于代表溶酶体区室的核周颗粒内的 HS-24、SB-3 和 Wi-38 成纤维细胞中。在 SB-3 细胞中，我们还以斑点分布定位了一小部分与质膜结合的酶，可以从外部接触到抗体。这种组织蛋白酶 B 分布的直接证明支持了有关该酶在肿瘤细胞中双重定位的生化数据。它还支持组织蛋白酶 B 直接参与细胞外基质降解的可能性，从而支持该酶在侵袭和转移中的贡献。
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 metastastis 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 lyosomal 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.