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Jolkinolide B, a bioactive diterpene isolated from the roots of Euphorbia fischeriana Steud, has various biological and pharmacological properties. In this study, the cytotoxicity of highly purifie...

Induction of apoptosis in K562 cells by jolkinolide B

BACKGROUND & OBJECTIVE Phosphorus plays a crucial role in metabolism. Some N-phosphoryl amino acids and N-phosphopeptides have important biological activities and medicinal value. The current study was designed to investigate the apoptosis of K562 cells induced by N-phosphoryl dipeptide methyl ester. METHOD Cell growth inhibition of K562 cells induced by 15 kinds of N-phosphoryl dipeptide methyl esters was analyzed by using MTT assay. The nuclei were stained by Hoechst 33,258 and the morphologic changes were observed. DNA agarose gel electrophoresis, double-staining, and flow cytomery were used to detect early apoptosis of K562 cells. RESULT (DIPP-L-Leu)2-L-Lys-OCH3 was the compound screened with MTT method that had better growth inhibiting activity with the IC50 of 22.66 mumol/L. Chromatin condensation and nuclear fragmentation were seen under fluorescence microscope in the cells treated with Hoechst 33,258. DNA agarose gel electrophoresis showed nuclear fragmentation (DNA ladder). Early apoptotic cells were also detected by flow cytometry. CONCLUSION These results suggest that(DIPP-L-Leu)2-L-Lys-OCH3 could induce the apoptosis of K562 cells.

[Apoptosis of K562 cell induced by N-phosphoryl dipeptide methyl ester].

Apoptosis as a mechanism of deleting cells from tissues plays an important role in physiological and varieties of pathological situations, especially cancer conditions. In order to search for tumor cells apoptosis inducers, the inhibition effects on K562 cells of N-phosphoryl dipeptide methyl esters were studied by MTT assays, and (DIPP-L-Leu)2-L-Lys-OCH3 was the compound which had the best activity. From the studies of the typical apoptotic morphologic changes, DNA agarose gel electrophoresis, and flow cytometry analysis, it could be concluded that (DIPP-L-Leu)2-L-Lys-OCH3 could induce apoptosis of K562 cells in a dose-dependent manner, and the IC50 was 22.66 μmol/L according to MTT assays.

Apoptosis induced by (DIPP-L-Leu)2-L-Lys-OCH3 in K562 cells

Phosphorus plays a crucial role in life chemistry.1 Some N-phosphoryl amino acids and N-phosphoryl peptides have important biological activities.2,3 Now we synthesize N-phosphoryl leucyl-lysine methyl ester ((DIPP-Leu)2-Lys-OCH3), a kind of N-phosphoryl branched peptide, that can induce apoptosis in K562 cell lines and its IC50 is 21.4 μg/ml. Morphologic changes can be observed under the optics microcopy, DNA fragmentation into 180 bps is detected by agarose gel electrophoresis (Figure 1): (A) marker (B) negative ctr. (C) positive ctr: cis-platinum (D-G) (DIPP-Leu)2-Lys-OMe of different concentrations: 50, 100, 200, 400 μg/ml and the “Ap” peak which is considered to be the marker of apoptosis can also be seen in flow cytometry (Figure 2): (A) untreated (B) treated with 100 μg/ml of (DIPP-Leu)2-Lys-OMe.

Apoptosis Induced by a Kind of N -Phosphoryl Branched Dipeptide

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[Apoptosis of K562 cell induced by N-phosphoryl dipeptide methyl ester].

Apoptosis induced by (DIPP-L-Leu)2-L-Lys-OCH3 in K562 cells

Apoptosis Induced by a Kind of N -Phosphoryl Branched Dipeptide