ZGDHu-1 and fludarabine have a synergistic effect on apoptosis of chronic lymphocytic leukemia cells

Qiu,Liannv; Liu,Jinlin; Wang,Zhenni; Hu,Weixiao; Huang,Qiang; Zhou,Yonglie

doi:10.3892/or.2015.4115

ZGDHu-1 and fludarabine have a synergistic effect on apoptosis of chronic lymphocytic leukemia cells

Authors:
- Liannv Qiu
- Jinlin Liu
- Zhenni Wang
- Weixiao Hu
- Qiang Huang
- Yonglie Zhou
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Affiliations: Department of Clinical Laboratory, Zhejiang Provincial People's Hospital, Hangzhou, Zhejiang 310004, P.R. China, College of Pharmaceutical Science, Zhejiang University of Technology, Hangzhou, Zhejiang 310004, P.R. China, Department of Hematology, Zhejiang Provincial People's Hospital, Hangzhou, Zhejiang 310004, P.R. China
Published online on: July 8, 2015 https://doi.org/10.3892/or.2015.4115
Pages: 1239-1248

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Abstract

Previously, it was demonstrated that the novel proteasome inhibitor N,N'-di-(m-methylphenyi)-3,6-dimethyl-1,4-dihydro-1,2,4,5-tetrazine-1,4-dicarboamide (ZGDHu‑1) possesses activity against chronic lymphocytic leukemia (CLL). In the present study, we attempted to assess whether this drug has a synergistic effect with fludarabine on the apoptosis of CLL cells. Annexin V/PI staining, mitochondrial membrane potential (ΔΨm) and reactive oxygen species (ROS) levels were examined by flow cytometry in short-term cell culture of blood cells from untreated newly diagnosed patients ex vivo. Expression of active caspase-3 and the Bcl-2/Bax ratio for determination of apoptosis were also investigated by flow cytometry and western blot analysis. Our results revealed that the ZGDHu-1 may induce the apoptosis of CLL cells through the mitochondrial pathway and its pro-apoptotic effect is CLL-specific, not affecting normal lymphocytes. Most importantly, a combination of ZGDHu-1 and a non-cytotoxic dose of fludarabine had a synergistic apoptotic effect. To some extent, caspase-3 activation may be involved in the mechanism of the ZGDHu-1 synergistic cytotoxic effect with fludarabine, as well as the cleavage of PARP, consequently leading to apoptosis. Notably, the rate of apoptosis caused by ZGDHu-1 alone or in combination with fludarabine was independent of prognostic markers of CLL disease such as ZAP-70 and CD38 expression or clinical Rai classification stage. In conclusion, ZGDHu-1 exhibited a significant synergistic effect with fludarabine to induce the apoptosis of CLL cells, which implies a possible clinical application.

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