N-(2-Hydroxypropyl)methacrylamide (HPMA) copolymer carrier containing the anticancer drug doxorubicin bound either by a proteolytically degradable bond (non-targeted PK1 or targeted with alpha-CD71 mAb) or by a hydrolytically degradable bond were synthesised and tested in vivo for various biological properties. Mouse 38C13 B-cell lympoma was used as a well established and defined cell line for this study. 38C13 cells are sensitive to free doxorubicin and IC50 was very low, about 0.014 microM. PK1 showed a strongly decreased cytostatic effect, IC50 being 12.6 microM. alpha-CD71 targeted conjugate, which can be considered as an antibody-targeted form of PK1, had IC50 0.358 microM. HPMA copolymer with doxorubicin bound via a hydrolytically sensitive bond (HYD conjugate) showed a high cytostatic effect with IC50 about 0.052 microM. We demonstrated that HYD conjugate inhibited DNA synthesis and induced p21(Waf1/Cip1) protein expression (p21(Waf1/Cip1) is cyclin-dependent kinase inhibitor which blocks cell cycle progression) as quickly as free doxorubicin, whereas PK1 acted much more slowly. Similarly, apoptosis induction measured by Annexin V binding and Caspase 3 activity was detected later after incubation of cells with PK1 or alpha-CD71 targeted conjugate. Apoptosis was manifested by elevation of bax and bad mRNA levels, which was much more rapid and intense in the case of free doxorubicin and HYD conjugate. Expression of antiapoptotic genes as well as cyclin-dependent kinases was surprisingly not affected.
- MeSH
- akrylamidy chemická syntéza metabolismus farmakologie MeSH
- apoptóza účinky léků MeSH
- buněčná membrána účinky léků patologie MeSH
- buněčný cyklus genetika MeSH
- DNA antagonisté a inhibitory genetika metabolismus MeSH
- doxorubicin chemická syntéza metabolismus farmakologie MeSH
- exprese genu účinky léků genetika MeSH
- geny myc účinky léků genetika MeSH
- hydrazony chemická syntéza metabolismus farmakologie MeSH
- hydrolýza MeSH
- inhibiční koncentrace 50 MeSH
- inhibitor p21 cyklin-dependentní kinasy MeSH
- kaspasa 3 MeSH
- kaspasy škodlivé účinky účinky léků metabolismus MeSH
- ligandy * MeSH
- messenger RNA genetika MeSH
- myši MeSH
- nádorové buněčné linie MeSH
- preklinické hodnocení léčiv metody MeSH
- proliferace buněk účinky léků MeSH
- protein Bad MeSH
- protein X asociovaný s bcl-2 MeSH
- proteiny buněčného cyklu genetika metabolismus MeSH
- protoonkogenní proteiny c-bcl-2 genetika metabolismus MeSH
- receptory transferinu účinky léků genetika MeSH
- thymidin metabolismus MeSH
- transportní proteiny genetika metabolismus MeSH
- tritium MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
- Názvy látek
- akrylamidy MeSH
- Bad protein, mouse MeSH Prohlížeč
- Bax protein, mouse MeSH Prohlížeč
- Casp3 protein, mouse MeSH Prohlížeč
- Cdkn1a protein, mouse MeSH Prohlížeč
- DNA MeSH
- doxorubicin MeSH
- hydrazony MeSH
- inhibitor p21 cyklin-dependentní kinasy MeSH
- kaspasa 3 MeSH
- kaspasy MeSH
- ligandy * MeSH
- messenger RNA MeSH
- N-(2-hydroxypropyl)methacrylamide co-polymer-doxorubicin conjugate MeSH Prohlížeč
- protein Bad MeSH
- protein X asociovaný s bcl-2 MeSH
- proteiny buněčného cyklu MeSH
- protoonkogenní proteiny c-bcl-2 MeSH
- receptory transferinu MeSH
- thymidin MeSH
- transportní proteiny MeSH
- tritium MeSH
