The antiproliferative and antitumor effect of wheat leaf ribonuclease was tested in vitro on the human ML-2 cell line and in vivo on athymic nude mice bearing human melanoma tumors. The antiproliferative activity of this plant ribonuclease was negligible in comparison with bovine seminal ribonuclease. In the experiments in vivo, a significant decrease of the tumor size, however, was observed in the mice treated with wheat leaf ribonuclease (27 kDa) compared with the control RNase A and polyethylene glycol. In nude mice injected intratumoraly with wheat leaf ribonuclease, the tumor size decreased from 100% in the control mice to 39% in treated mice. In the mice treated with polyethylene glycol-conjugated wheat leaf ribonuclease, the tumor reduction was observed from 100 to 28%, whereas in counterparts treated with polyethylene glycol-conjugated bovine seminal ribonuclease the tumor inhibition was reduced from 100 to 33%. Certain aspermatogenic and embryotoxic activity of wheat leaf ribonuclease and bovine seminal ribonuclease also appeared, but was lower in comparison with the effect of onconase. Mutual immunological cross-reactivity between wheat leaf ribonuclease antigens on one side and animal RNases (bovine seminal ribonuclease, RNase A, human HP-RNase and onconase) on the other side proved a certain structural similarity between animal and plant ribonucleases. Immunogenicity of wheat leaf ribonuclease was weaker in comparison with bovine seminal ribonuclease (titer of antibodies 160-320 against 1280-2560 in bovine seminal ribonuclease). Interestingly, immunosuppressive effect of wheat leaf ribonuclease tested on mixed lymphocyte culture-stimulated human lymphocytes reached the same level as that of bovine seminal RNase. The antibodies against wheat leaf ribonuclease produced in the injected mice did not inactivate the biological effect of this plant RNase in vivo. This is probably the first paper in which plant ribonuclease was used as antiproliferative and antitumor drug against animal and human normal and tumor cells and tissues in comparison with animal ribonucleases.
- MeSH
- antitumorózní látky fytogenní izolace a purifikace terapeutické užití toxicita MeSH
- embryo savčí účinky léků MeSH
- financování organizované MeSH
- imunosupresiva MeSH
- injekce intraperitoneální MeSH
- lidé MeSH
- listy rostlin chemie MeSH
- lymfocyty imunologie účinky léků MeSH
- myši nahé MeSH
- myši MeSH
- nádorové buněčné linie MeSH
- nádory farmakoterapie terapie MeSH
- polyethylenglykoly MeSH
- proliferace buněk účinky léků MeSH
- pšenice enzymologie MeSH
- ribonukleasy MeSH
- spermatogeneze účinky léků MeSH
- těhotenství MeSH
- testis patologie účinky léků MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- myši MeSH
- těhotenství MeSH
- ženské pohlaví MeSH
- zvířata MeSH
Previously we have shown that monomeric RNase A has no significant biological activity, whereas its oligomers (dimer to tetramer) prepared by lyophilizing from 50% acetic acid solutions, show remarkable aspermatogenic and antitumor activities. Furthermore, conjugates prepared by chemical binding of native RNase A to polyethylene glycol (PEG) have shown a significant aspermatogenic and antitumor activities. In this work we show that the chemical conjugation of PEG to the RNase A C-dimer, and to the two RNase A trimers (NC-trimer and C- trimer) decreases the aspermatogenic activity of the oligomers while increasing their inhibitory activity on the growth of the human UB900518 amelanotic melanoma transplanted in athymic nude mice. Moreover, the PEG-conjugated RNaseA oligomers are devoid, like the free oligomers, of any embryotoxic activity.
- MeSH
- antispermatogenní látky farmakologie MeSH
- antitumorózní látky farmakologie chemie MeSH
- dimerizace MeSH
- embryo savčí účinky léků MeSH
- lidé MeSH
- melanom experimentální farmakoterapie MeSH
- myši MeSH
- nádorové buněčné linie MeSH
- pankreatická ribonukleasa farmakologie chemie MeSH
- peptidové fragmenty farmakologie chemie MeSH
- polyethylenglykoly farmakologie chemie MeSH
- spermatogeneze účinky léků MeSH
- transplantace nádorů MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- práce podpořená grantem MeSH
DNA methylation/demethylation pattern, determined by 5-methylcytosine (5-MeC) immunostaining, was evaluated in porcine "in vivo" produced embryos from zygote up to the blastocyst stage. In one-cell stage embryos, only the maternal pronucleus showed a positive labeling whilst the paternal pronucleus showed almost no labeling. The intensity of labeling is high until the late morula stage. Blastocysts containing less than 100 cells showed the same intensity of labeling in both the inner cell mass (ICM) nuclei and the trophectodermal (TE) cell nuclei. Interestingly, with further cell multiplication, cells of the ICM became more intensively labeled when compared to TE cells. This distinct methylation pattern is even more profound in blastocysts containing about 200-300 cells and is not caused by the difference in the cell volume of ICM and TE cells.
- MeSH
- antispermatogenní látky farmakologie MeSH
- antitumorózní látky farmakologie MeSH
- embryo savčí účinky léků MeSH
- endonukleasy specifické pro jednořetězcové nukleové kyseliny * farmakologie imunologie MeSH
- experimentální nádory * farmakoterapie MeSH
- fazol * enzymologie MeSH
- lidé MeSH
- myši nahé MeSH
- myši MeSH
- nádorové buněčné linie MeSH
- pankreatická ribonukleasa farmakologie imunologie MeSH
- proliferace buněk účinky léků MeSH
- spermatogeneze účinky léků MeSH
- spermie účinky léků MeSH
- teratogeny farmakologie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
