组织蛋白酶L及其内源性抑制剂在永生化和转化成纤维细胞中的表达
Expression of cathepsin L and its endogenous inhibitors in immortal and transformed fibroblasts.
作者信息
Dilakyan E A, Zhurbitskaya V A, Vinokurova S V, Gureeva T A, Lubkova O N, Topol L Z, Kisseljov F L, Solovyeva N I
机构信息
Institute of Biomedical Chemistry, Russian Academy of Medical Sciences (RAMS), Pogodinskaya St. 10, 119992, Moscow, Russia.
出版信息
Clin Chim Acta. 2001 Jul 5;309(1):37-43. doi: 10.1016/s0009-8981(01)00495-8.
METHODS
The activities of cathepsin L and its endogenous inhibitors were analyzed in rat embryo fibroblasts, immortalized and transformed by different genes.
RESULTS
Regardless of the transfecting agent used (DNA of adenovirus SA7 or polyomavirus LT gene), the immortal cells showed an increase in the cathepsin L activity (in both lysates and conditioned media) compared to primary fibroblasts. Transformed cells exhibited either an increase (with c-Ha-ras gene) or decrease (with E7 HPV gene) in cathepsin L activity in lysates as opposed to immortal cells.
CONCLUSIONS
The data are suggestive of alterations in the trafficking of cathepsin L upon fibroblast transfection with polyomavirus LT gene and E7 HPV gene. An endogenous inhibitor(s) of cysteine proteinase was found in conditioned media, but not in lysates, of all cell cultures studied and its activity in normal fibroblasts was higher than in media of immortal and transformed cells.
方法
在通过不同基因永生化和转化的大鼠胚胎成纤维细胞中分析组织蛋白酶L及其内源性抑制剂的活性。
结果
无论使用何种转染剂（腺病毒SA7的DNA或多瘤病毒LT基因），与原代成纤维细胞相比，永生化细胞的组织蛋白酶L活性（在裂解物和条件培养基中）均有所增加。与永生化细胞相反，转化细胞的裂解物中组织蛋白酶L活性要么增加（用c-Ha-ras基因），要么降低（用E7 HPV基因）。
结论
数据表明，在用多瘤病毒LT基因和E7 HPV基因转染成纤维细胞后，组织蛋白酶L的运输发生了改变。在所有研究的细胞培养物的条件培养基中发现了一种半胱氨酸蛋白酶的内源性抑制剂，但在裂解物中未发现，并且其在正常成纤维细胞中的活性高于永生化细胞和转化细胞的培养基中的活性。
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