Avtor/Urednik		Premzl, Aleš; Kos, Janko
Naslov		Cysteine and aspartic proteases cathepsins B and D determine the invasiveness of MCF10A neoT cells
Prevedeni naslov		Cisteinski in aspartatni proteazi katepsina B in D določata invazivnost MCF10A neoT celic
Tip		članek
Vir		Radiol Oncol
Vol. in št.		Letnik 37, št. 4
Leto izdaje		2003
Obseg		str. 241-8
Jezik		eng
Abstrakt		Background. Lysosomal cathepsins B and D have been reported to play a role in various processes leading to progression of malignant disease. In ras-transformed MCF10A neoT cells both enzymes show similar vesicular distribution in perinuclear and peripheral cytoplasmic regions. Results. The co-localization of cathepsins B and D in some vesicles as defined by confocal microscopy supports their co-ordinate activity in the proteolytic cascade. On the other hand, we showed that stefin A, an endogenous intracellular inhibitor of cysteine proteases, did not co-localize with cathepsin B and is presumably not involved in regulation of its enzymatic activity within the vesicles. Intracellular localization of both enzymes was confined to similar vesicles as the fluorescent degradation products of DQ-collagen Iv either in individual cells or cell spheroids. The capability of these two enzymes to degrade collagen and other components of extracellular matrix is further supported by the results of Matrigel invasion assay. We showed that specific intracellular (CA-074 Me) and extracellular (CA-074) inhibitors of cathepsin B and pepstatin A, an inhibitor of cathepsin D, significantly reduced invasion of MCF10A neoT cells. Our results also show that in contrast to some other studies the activation peptide of pro-cathepsin D exhibited no mitogenic effect on MCF10A neoT, MCF-7 or HEK 293 cells. Conclusion. We conclude that lysosomal eysteine proteases eathepsins B and D predominantly participate in degradation of extracellular matrix and facilitate invasion of tumour cells.
Izvleček		Izhodišča. Znano je, da imata lizosomska katepsina B in D pomembno vlogo v različnih procesih, ki vodijo do napredovanja malignih bolezni. V ras-transformiranih MCF10A neoT celicah oba encima kažeta podobno vezikularno razporeditev, tako v perinuklearnih kot v perifernih citoplazemskih regijah. Rezultati. V nekaterih veziklih je bila s pomočjo konfokalne mikroskopije določena njuna kolokalizacija, kar potrjuje odvisno delovanje v proteolitski kaskadi. Za stefin A, endogeni znotrajcelični inhibitor cisteinskih proteaz, pa smo pokazali da se ne kolokalizira s katepsinom B in predvidoma ni udeležen pri regulaciji njegove aktivnosti znotraj veziklov. Znotrajcelična lokalizacija obeh katepsinov se ujema z lokalizacijo razgradnih produktov DQ-kolagena IV, bodisi v posameznih celicah ali v celičnih sferoidih. Sposobnost katepsinov B in D, da razgrajujeta kolagen in druge komponente zunajceličnega matriksa, potrjujejo tudi rezultati testa razgradnje matrigela. Zaključki. Pokazali smo, da specifična inhibitorja katepsina B (znotrajcelični CA-074 Me in zunajcelični CA-074) in pepstatin A, inhibitor katepsina D, značilno zmanjšajo invazijo MCF10A neoT celic. Naši rezultati tudi kažejo, da v nasprotju z nekaterimi prejšnjimi študijami aktivacijski peptid pro-katepsina D ni mitogen na MCF10A neoT, MCF-7 in HEK-293 celice.
Deskriptorji		TUMOR CELLS, CULTURED CATHEPSIN B CATHEPSIN D NEOPLASM INVASIVENESS IMMUNOHISTOCHEMISTRY