Induction of apoptosis by osthole in HL-60 cells and the molecular mechanism research

Abstract

Abstract:Objective To detect the effect of osthole on proliferation and apoptosis of HL-60 cells and its molecular mechanism. Methods HL-60 cells proliferation was measured through the CCK8 assay method. The cell morphological changes were observed by Hoechst33342 staining after 8 h of drug effect. Induction of apoptosis was determined by flow cytometry and fluorescent microscopy. Expressions of Bcl-2 and Bax mRNA were evaluated by RT-PCR, and the expressions of cleaved caspase-3, caspase-8, caspase-9, Fas and FasL were evaluated by using western bolt assay. Results Osthole could inhibit the proliferation of HL-60 cells, the maximum inhibiting rate was （90.7±4.5）%， F=138.46, P=0.000; the apoptosis rate was 33.6%， F=27.75, P=0.006. The changes of apoptosis of cells and nucleus were shown in cell morphological observation. Osthole affected the decrease of the mRNA levels of Bcl-2 and the increase of the Bax mRNA levels via a dosedependent manner（F=210.12, P=0.000）. Western blotting demonstrated that osthole could lead to the increase of the expression levels of cleaved caspase-3, caspase-8, caspase-9, Fas and FasL in the HL-60 cell line via a timedependent manner. Conclusion Data suggests that osthole inhibits proliferation and induces apoptosis of HL-60 cells through mitochondriadependent pathway and deathreceptor pathway.

Key words:Osthole,Leukemia,Apoptosis

Wang Yan, Jiang Guosheng, Ren Xia, Huang Ning, Bi Kehong. Induction of apoptosis by osthole in HL-60 cells and the molecular mechanism research[J]. Journal of International Oncology, 2014, 41(5): 371-375.

References

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