Identification of heplipin inducing apoptosis-related genes in KG-1 human leukemia cells by differential display RT-PCR / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate whether Heplipin can induce KG-1 cell apoptosis and explore apoptosis related differentially expressed genes in KG-1 leukemia cell before and after Heplipin induction.</p><p><b>METHODS</b>DNA distribution and DNA electrophoresis were used to prove that Heplipin can induce KG-1 cell apoptosis. The differential display reverse transcription-polymerase chain reaction (DDRT-PCR) was adopted to screen differentially expressed genes before and after Heplipin induction of KG-1 cells for 16 hours and 20 hours. The differentially expressed genes were cloned and analyzed.</p><p><b>RESULTS</b>Heplipin could induce KG-1 cell apoptosis. There were differentially expressed genes in KG-1 cells before and after induction. Wnt13 and ATPase 3 were apoptosis related differentially downregulated genes after Heplipin induction. Conclusion Heplipin can induce KG-1 cell apoptosis. Heplipin induced KG-1 cell apoptosis is related with Wntl3 and ATPase3 (PSMC3). It is the first report that Wnt13 was detected in leukemia cell line.</p>

In vitro cultivation of dendritic cells with serum-free medium / 中国实验血液学杂志

This study was aimed to investigate the protocol in vitro to incubate the dendritic cell (DC) derived from peripheral blood monocytes using serum-free medium X-VIVO 20. Peripheral blood monocytes from healthy donors were treated with 100 ng/ml GM-CSF and 500 U/ml IL-4, respectively. After cultivation for 6 days, they were treated with 100 ng/ml calcium ionophore A23187. After cultivation for 24 hours the cellular morphology was observed under invert microscope, the surface markers were analyzed by flow cytometry, the proliferation of allogenetic T cells was detected by MTT colorimetry, the specific cytotoxicity of T cells primed with DC was examined by MTT assay. The results showed that in all three groups with serum-free, fetal calf serum (FCS) and human AB serum mediums, cells displayed characteristic morphological features of DC. Simultaneously CD14 expression was decreased, and CD83, HLA-DR and CDw123 expression were increased on these cells. In addition, DCs cultured with these methods could evidently stimulate the proliferation of allogenetic T cell. As compared with the two controls of serum containing groups, the cultured cells in the serum-free groups showed almost the same allo-stimulatory capability and cellular morphology and surface markers, and T lymphocytes primed with the culture-derived DC exhibited the similar killing activity to K562 (P > 0.05). It is concluded that there is no significance in DC numbers, morphology, epitope and ability to stimulate the proliferation of allogenetic T cells between DC induced by serum-free X-VIVO 20 medium and DC induced by serum-contained medium. DC cultured and induced by serum-free medium is worth using in practice widely.