Effects of N, N-di-(m-methylphenyl)-3, 6-dimethyl-1, 4-dihydro-1,2,4, 5-tetrazine-1,4-dicarboxamide (ZGDhu-1) on SHI-1 leukemia cells in vitro / 中华血液学杂志

ABSTRACT

<p><b>OBJECTIVE</b>To study the effect of ZGDHu-1 on proliferation, differentiation and apoptosis in SHI-1 human leukemia cell line and explore its possible mechanism. Methods SHI-1 cells were cultured with different concentration of ZGDHu-1 and for different time. The cell proliferation was analysed by cell counting, alive cell count, MTT assay and Brdu-ELISA. Cell apoptosis was analysed by morphology, DNA content, Annexin-V/PI and Hoechst 33258 labeling method. Cell differentiation were assayed by morphology,expression of CD11b,CD14 and CD64 and NBT reduction. The expressions of phosphorylated p38MAPK or STAT3 were analysed by flow cytometry.</p><p><b>RESULTS</b>ZGDHu-1 inhibited SHI-1 cell proliferation in a time and dose dependent manner, the IC50- 48 h and IC50- 72 h were 250 ng/ml and 85 ng/ml, respectively. The majority of SHI-1 cells were arrested in G2/M phase. 48h after treated with 200 ng/ml ZGDHu-1, and those in G2/M phase accounted for (48.4 +/- 2.1)%. The SHI-1 cells apoptosis was increased with a time- and does-dependent manner. The morphology of SHI-1 cells cultured with 2-50 ng/ml ZGDHu-1 for three days become more mature with higher NBT positivity and up-regulated expressions of CD11b,CD14 and CD64. The expression of phosphor-p38MAPK was increased and phosphor-STAT3 down-regulated by the treatment of ZGDHu-1.</p><p><b>CONCLUSION</b>ZGDHu-1 can inhibit SHI-1 cell proliferation and induce the cell differentiation and apoptosis. The mechanism may associate with its up-regulation of phosphor-p38MAPK and down-regulation phosphor-STAT3.</p>