Breast cancer cells express cathepsins B and L but not cathepsins K or H.

Cancer biochemistry biophysics Pub Date : 1999-07-01
O Ishibashi, Y Mori, T Kurokawa, M Kumegawa
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Abstract

Lysosomal cysteine proteinases (cathepsins) are considered to play a role in bone degradation mediated by metastatic breast cancers. To evaluate which cathepsin contributes to the osteolysis, we quantitatively determined the expression levels of four cathepsins in two breast cancer cell lines, MCF-7 and MDA-MB-231, by competitive RT-PCR. Cathepsin K, which is the most abundant cathepsin in osteoclasts, was not detected in either cell lines. We also failed to detect cathepsin H mRNA. By contrast, we found significant expression of cathepsins B and L in both cell lines. By Northern blot analysis cathepsin B mRNA was detected in a single form in these cells, whereas osteoclasts contained multiple forms of the mRNA. Cathepsin B protein was also detected by Western blotting as a single immunoreactive band corresponding to its mature enzyme. These findings suggest that osteolysis associated with metastatic breast cancers takes place in a different way from osteoclast-mediated bone resorption.

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乳腺癌细胞表达组织蛋白酶B和L,但不表达组织蛋白酶K和H。
溶酶体半胱氨酸蛋白酶(组织蛋白酶)被认为在转移性乳腺癌介导的骨降解中起作用。为了评估哪种组织蛋白酶有助于骨溶解,我们通过竞争性RT-PCR定量测定了两种乳腺癌细胞系MCF-7和MDA-MB-231中四种组织蛋白酶的表达水平。组织蛋白酶K是破骨细胞中最丰富的组织蛋白酶,在两种细胞系中均未检测到。我们也没有检测到组织蛋白酶H mRNA。相比之下,我们发现组织蛋白酶B和L在两种细胞系中都有显著表达。通过Northern blot分析,在这些细胞中检测到组织蛋白酶B mRNA的单一形式,而破骨细胞中则含有多种形式的mRNA。组织蛋白酶B蛋白也通过Western blotting检测为其成熟酶对应的单一免疫反应带。这些发现表明,与转移性乳腺癌相关的骨溶解发生的方式与破骨细胞介导的骨吸收不同。
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