Many conjugates of water-soluble polymers with biologically active molecules were developed during the last two decades. Although, therapeutic effects of these conjugates are affected by the properties of carriers, the properties of the attached drugs appear more important than the same carrier polymer in this case. Pirarubicin (THP), a tetrahydropyranyl derivative of doxorubicin (DOX), demonstrated more rapid cellular internalization and potent cytotoxicity than DOX. Here, we conjugated the THP or DOX to N-(2-hydroxypropyl)methacrylamide copolymer via a hydrazone bond. The polymeric prodrug conjugates, P-THP and P-DOX, respectively, had comparable hydrodynamic sizes and drug loading. Compared with P-DOX, P-THP showed approximately 10 times greater cellular uptake during a 240 min incubation and a cytotoxicity that was more than 10 times higher during a 72-h incubation. A marginal difference was seen in P-THP and P-DOX accumulation in the liver and kidney at 6 h after drug administration, but no significant difference occurred in the tumor drug concentration during 6-24 h after drug administration. Antitumor activity against xenograft human pancreatic tumor (SUIT2) in mice was greater for P-THP than for P-DOX. To sum up, the present study compared the biological behavior of two different drugs, each attached to an N-(2-hydroxypropyl)methacrylamide copolymer carrier, with regard to their uptake by tumor cells, body distribution, accumulation in tumors, cytotoxicity, and antitumor activity in vitro and in vivo. No differences in the tumor cell uptake of the polymer-drug conjugates, P-THP and P-DOX, were observed. In contrast, the intracellular uptake of free THP liberated from the P-THP was 25-30 times higher than that of DOX liberated from P-DOX. This finding indicates that proper selection of the carrier, and especially conjugated active pharmaceutical ingredient (API) are most critical for anticancer activity of the polymer-drug conjugates. THP, in this respect, was found to be a more preferable API for polymer conjugation than DOX. Hence the treatment based on enhanced permeability and retention (EPR) effect that targets more selectively to solid tumors can be best achieved with THP, although both polymer conjugates of DOX and THP exhibited the EPR effects and drug release profiles in acidic pH similarly.
- MeSH
- akrylamidy chemie MeSH
- antibiotika antitumorózní chemie farmakologie MeSH
- antitumorózní látky chemie farmakologie MeSH
- doxorubicin analogy a deriváty chemie farmakologie MeSH
- experimentální sarkom farmakoterapie metabolismus patologie MeSH
- lidé MeSH
- myši inbrední BALB C MeSH
- myši MeSH
- nádorové buňky kultivované MeSH
- nosiče léků aplikace a dávkování chemie MeSH
- polymery aplikace a dávkování chemie MeSH
- proliferace buněk účinky léků MeSH
- xenogenní modely - testy antitumorózní aktivity MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
Tumor models are essential for basic anticancer research and development of novel therapies. In this study, we used a rat sarcoma model in which subcutaneous tumor develops after D6 cell inoculation. The aim of the current study was to analyze changes in haematological parameters, immune cell sub-populations and cytokine profiling during tumor growth, after tumor excision and after second inoculation of D6 cells. Tumor progression was found to be associated with an increased number of leukocytes and increased proportion of CD11b+ cells in peripheral blood. Serum concentration of chemokine (c-c motif) ligand 2, L-selectin and intra cellular adhesion molecule-1 also increased with growing tumor. However, the proportion of CD4+, CD8+ and MHC II+ cells decreased with growth of tumors. After tumor excision, all these parameters returned to pre-inoculation levels and did not change even after a second inoculation of D6 cells. Moreover, absence of secondary tumors after second inoculation of D6 cells gives an insight into development of antitumor immunity stimulated by primary tumor.
- MeSH
- antigeny CD11b krev MeSH
- chemokin CCL2 krev MeSH
- experimentální sarkom krev patologie MeSH
- krysa rodu rattus MeSH
- leukocytóza MeSH
- potkani inbrední LEW MeSH
- progrese nemoci MeSH
- průtoková cytometrie MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
UNLABELLED: Comparing the gene expression profiles of metastatic and nonmetastatic cells has the power to reveal candidate metastasis-associated genes, whose involvement in metastasis can be experimentally tested. In this study, differentially expressed genes were explored in the v-src-transformed metastatic cell line PR9692 and its nonmetastatic subclone PR9692-E9. First, the contribution of homeodomain only protein X (HOPX) in metastasis formation and development was assessed. HOPX-specific knockdown decreased HOPX expression in the nonmetastatic subclone and displayed reduced cell motility in vitro. Critically, HOPX knockdown decreased the in vivo metastatic capacity in a syngeneic animal model system. Genomic analyses identified a cadre of genes affected by HOPX knockdown that intersected significantly with genes previously found to be differentially expressed in metastatic versus nonmetastatic cells. Furthermore, 232 genes were found in both screens with at least a two-fold change in gene expression, and a number of high-confidence targets were validated for differential expression. Importantly, significant changes were demonstrated in the protein expression level of three metastatic-associated genes (NCAM, FOXG1, and ITGA4), and knockdown of one of the identified HOPX-regulated metastatic genes, ITGA4, showed marked inhibition of cell motility and metastasis formation. These data demonstrate that HOPX is a metastasis-associated gene and that its knockdown decreases the metastatic activity of v-src-transformed cells through altered gene expression patterns. IMPLICATIONS: This study provides new mechanistic insight into a HOPX-regulated metastatic dissemination signature.
- MeSH
- buněčný cyklus MeSH
- down regulace MeSH
- experimentální sarkom genetika patologie sekundární MeSH
- forkhead transkripční faktory genetika metabolismus MeSH
- genový knockdown MeSH
- geny src MeSH
- homeodoménové proteiny genetika metabolismus MeSH
- kur domácí MeSH
- metastázy nádorů genetika MeSH
- molekuly buněčné adheze nervové genetika metabolismus MeSH
- nádorová transformace buněk genetika MeSH
- nádorové buněčné linie MeSH
- pohyb buněk MeSH
- ptačí proteiny genetika metabolismus MeSH
- regulace genové exprese u nádorů MeSH
- sekvenční analýza hybridizací s uspořádaným souborem oligonukleotidů MeSH
- stanovení celkové genové exprese MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Experiments were designed to assess age-related changes in the transplantability of human tumours xenografted in congenitally athymic (nu/nu) mice. It has been found that the number of progressively growing human tumour xenografts decreased significantly with increasing age of BALB/c nu/nu recipients. These findings, taken together with a previously recognized increase in the frequency of endogenous interleukin 2 (IL-2)-producing cells with age of nu/nu mice, prompted us to investigate whether administration of exogenous IL-2 to young adult nu/nu mice could change the transplantability of human tumours in the mice. Peritumoral administration of exogenous interleukin 2 to 8-week-old nu/nu mice inhibited the growth of the human tumour xenografts. In vitro activation of nu/nu splenocytes with exogenous Il-2 resulted in the generation of killer cells which have been found to be cytolytic when allowed to react with human tumour targets in 51Cr cytotoxicity assay. In addition, it has been found that the percentage of IL-2-activated Thy 1.2+ and ASGM1+ cells substantially increased with increasing age of nu/nu spleen cell donors. These findings are compatible with the hypothesis that the observed age-related decrease in takes of human tumour xenografts might be determined by the increasing level of IL-2 production and subsequent maturation of IL-2-dependent effector cells.
- MeSH
- antigeny povrchové analýza MeSH
- buněčný cyklus MeSH
- buňky K aktivované lymfokiny imunologie MeSH
- cytotoxicita imunologická MeSH
- experimentální sarkom patologie MeSH
- HeLa buňky MeSH
- interleukin-2 farmakologie MeSH
- karcinom z přechodných buněk patologie MeSH
- lidé MeSH
- lymfom T-buněčný patologie MeSH
- myši nahé růst a vývoj MeSH
- myši MeSH
- nádory děložního čípku patologie MeSH
- nádory močového měchýře patologie MeSH
- průtoková cytometrie MeSH
- slezina imunologie růst a vývoj MeSH
- stárnutí fyziologie MeSH
- transplantace heterologní MeSH
- transplantace nádorů MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- MeSH
- ascites patologie MeSH
- ascitická tekutina MeSH
- buněčné dělení MeSH
- chromozomální aberace MeSH
- chromozomy MeSH
- diploidie MeSH
- experimentální sarkom patologie MeSH
- homologní transplantace MeSH
- karyotypizace MeSH
- myši MeSH
- nehodgkinský lymfom patologie MeSH
- polyploidie MeSH
- transplantace nádorů MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- myši MeSH
- zvířata MeSH
- MeSH
- antigeny analýza MeSH
- antisérum analýza MeSH
- erytrocyty chemie imunologie MeSH
- experimentální sarkom chemicky indukované imunologie krev patologie MeSH
- krysa rodu rattus MeSH
- kultivační techniky MeSH
- peritoneum imunologie patologie MeSH
- železo-dextranový komplex MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- zvířata MeSH
