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Molecular cloning and sequencing of partial cDNA of tumor necrosis factor and p75 tumor necrosis factor receptor of Syrian golden hamster (Mesocricetus auratus) with the use of universal primers.

100%

Dobrzańska Z., Więckiewicz J., Bigda J.

2002

vol. 49

issue 2

427-431

In this study we cloned and analysed partial cDNA of tumor necrosis factor (TNF) and p75 TNF-R receptor of Syrian golden hamster (Mesocricetus auratus). We obtained a 382-bp sequence of TNF and a 148-bp sequence coding for p75 TNF-R. The primers used for the cloning were designed on the basis of inter-species homology, thus presumably can be used for cloning and analysis of TNF and p75 TNF-R genes of other mammals.

Antitumor activity of antimicrobial peptides against U937 histiocytic cell line

88%

Koszałka P., Kamysz E., Wejda M., Kamysz W., Bigda J.

We investigated cytotoxic activity of antimicrobial peptides of different origin (both naturally occurring and synthetic), structure and known mechanisms of action against human histiocytic lymphoma cell line U937. The strongest cytotoxic activity against U937 cell line was shown by Pexiganan MSI-78, followed by Citropin 1.1, Protegrin 1 and a synthetic lipopeptide, N-α-palmitoyl-l-lysyl-l-lysine amide (Pal-Lys-Lys-NH2). The cytotoxic activity of the peptides was more dependent on the time of incubation than concentration. Only for the lipopeptide, whose mode of action was restricted to disruption of electric potential of the cell membrane, the correlation between cytotoxicity and concentration was almost linear. The high cytotoxicity of Pexiganan MSI-78, Protegrin 1 and the lipopeptide could be basically explained by their membranolytic activity leading to necrosis. However, in the case of Citropin 1.1, the cell membrane integrity was disrupted only slightly and independently of the peptide concentration. Therefore, some other mechanism of action might be responsible for its strong dose-dependent cytotoxic activity, e.g., membranolytic activity leading to apoptosis. Furthermore, TNF-α production due to LPS (lipopolysaccharide) stimulation was suppressed by the presence of Citropin 1.1, Pexiganan MSI-78 or Protegrin 1, but not by Buforin 2 or the lipopeptide. Our experiments have shown that cytotoxic activity is not limited to some specific molecular structure of a peptide, but rather to the length of the peptide chain as it is likely to affect the efficiency of the tumor cell membrane disruption and interaction with LPS.

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2 Acta Biochimica Polonica

1 2011

1 2002

2 Bigda J.

1 Dobrzańska Z.

1 Kamysz E.

1 Kamysz W.

1 Koszałka P.

1 Wejda M.

1 Więckiewicz J.

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Molecular cloning and sequencing of partial cDNA of tumor necrosis factor and p75 tumor necrosis factor receptor of Syrian golden hamster (Mesocricetus auratus) with the use of universal primers.

Antitumor activity of antimicrobial peptides against U937 histiocytic cell line