Roles and mechanisms of Fas/FasL in the apoptosis of HL-60 cells induced by euphorbiasteroid

doi:10.3760/cma.j.issn.1673422X.2014.09.012

Abstract

Abstract: ObjectiveTo investigate the effect of euphorbiasteroid on inducing the apoptosis of HL60 cells and demonstrate whether the Fas/FasL signaling pathway is involved in the induction of apoptosis. MethodsHL60 cells were treated with dose of 2.5, 10, 40 μg/ml of euphorbiasteroid in vitro for 24 h respectively. After that, cell counting Kit8 was used to detect cell proliferation. The morphology of HL60 cells were observed under light and fluorescent microscopy. The early cell apoptosis was detected by using flow cytometry with Annexin ⅤFITC /PI double staining. The expressions of Fas, FasL, caspase8 and caspase3 mRNA were analyzed by the method of RTPCR. The activities of caspase8 and caspase3 were examined by chromatometry. ResultsCompared with 1640 control group, HL60 cell proliferation was inhibited significantly by euphorbiasteroid. The inhibition rates were (34.9±3.7)%, (54.6±5.2)% and (61.3±4.3)% respectively. Moreover, HL60 cells exhibited typical morphological features. Early cell apoptosis rates of HL60 cells were (23.4±3.1)%, (35.7±4.3)% and (53.2±3.9)% respectively. Furthermore, the expressions of Fas, FasL, caspase3 and caspase8 mRNA were upregulated significantly after euphorbiasteroid administration in a dosedependent manner (P＜0.01). After treated with euphorbiasteroid, the activities of caspase8 and caspase3 were significantly enhanced (P＜0.01). ConclusionThe upregulation effect of euphorbiasteroid on Fas/FasL signaling pathway might contribute to the apoptosis of HL60 cells in a dosedependent manner.

Key words: HL-60 cells, Apoptosis, Fas/FasL, Euphorbiasteroid

Guo Fei, Li Xia, Zhang Chao, Ren Xia, Shi Meiyan, Jiang Guosheng. Roles and mechanisms of Fas/FasL in the apoptosis of HL-60 cells induced by euphorbiasteroid[J]. Journal of International Oncology, 2014, 41(8): 679-684.

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