GM-CSF,IL-<em>3</em>和GM-CSF/IL-<em>3</em>融合蛋白对辐射致髓样白血病细胞株Tf-<em>1</em>凋亡的影响

• 作者: Li WEN Qin-yan GONG Ying-qing LU 于榕 姚明辉 杨素荣
• 来源期刊: 中国药理学报（英文版）
• 学科: 医学
• 关键词: granulocyte-macrophage colony-stimulating factor interleukin-3 recombinant fusion proteins cultured cells radiation apoptosis caspases flow cytometry
• 年，卷（期）: 2004,（1）
• 所属期刊栏目: 研究原著
• 页码范围: 68-75
• 页数: 8页
• 分类号: R733
• 语种: 英文

摘要

AIM: To observe the effects of three cytokines on the apoptosis of Tf- 1 cells induced by γ irradiation and investigate the relationship between apoptosis and caspase-3 activity. METHODS: Different cytokines GM-CSF, IL-3 and GM-CS/IL-3 fusion protein were added into the irradiated Tf-1 cells. MTT assay, morphology, flow cytometry,and DNA fragmentation assay were used to observe the effects of cytokines on apoptosis. The caspase-3 activity was determined with a fluorocytometer. RESULTS: Irradiated Tf-1 cells showed typical morphological characteristic of apoptosis demonstrated by transmission electron microscopy and were accumulated in G0/G1 phase. In the groups treated with growth factors after irradiation, three cytokines significantly increased the viability rate, distinctly decreased the apoptosis rate and the proportion of DNA fragmentation. When Tf- 1 cells were irradiated by γ irradiation, caspase-3 activity was increased at different time points. In comparison with the control group in which no growth factor was added after the cells were irradiated, the caspase-3 activity of irradiated Tf-1 cells was significantly inhibited by addition of the above cytokines. Thirty-six hours after irradiation, in the control group,GM-CSF, IL-3, GM-CSF and IL-3 in combination, and two GM-CSF/IL-3 fusion protein groups, the apoptosis rate was 73 %, 11%, 15 %, 13 %, 12 %, and 13 %. The percent of fragmented DNA was 36 %, 19 %, 18 %, 14 %,13 %, and 14 %. The fluorescence intensity was 16923, 5529, 6581, 5322, 5426, and 5485. CONCLUSION:GM-CSF, IL-3, and GM-CSF/IL-3 fusion protein could protect Tf-1 cells from apoptosis induced by γ irradiation.After Tf-1 cells were irradiated, the caspase-3 activity was significantly increased but was dramatically decreasedby the above cytokines. The remarkable inhibition of caspase-3 activity may be one of the mechanisms of thesehematopoietic growth factors exerting their anti-apoptotic effects.