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Molecular cloning of the Bombus terrestris bumblebee venom protein phospholipase A2 and its anti-leukemia effects on K562 cells
被引：2
|作者：
Qiu, Yuling
[1
]
Chen, Yali
[1
]
Yu, Haiyang
[2
]
Zhou, Qianxiang
[1
]
Wang, Ran
[1
]
Jin, Meihua
[1
]
Kong, Dexin
[1
,3
]
机构：
[1] Tianjin Med Univ, Sch Pharm, Tianjin Key Lab Technol Enabling Dev Clin Therape, Tianjin 300070, Peoples R China
[2] Tianjin Univ Tradit Chinese Med, Tianjin State Key Lab Modern Chinese Med, Tianjin 300193, Peoples R China
[3] Tianjin Med Univ, Res Ctr Basic Med Sci, Tianjin 300070, Peoples R China
基金：
中国国家自然科学基金;
中国博士后科学基金;
关键词：
PLA2;
Bumblebee venom;
Purification;
Anti-leukemia;
Apoptosis;
BEE VENOM;
IN-VITRO;
CANCER-THERAPY;
ERYTHROID-DIFFERENTIATION;
SERINE-PROTEASE;
STELLETTIN B;
WASP VENOMS;
APOPTOSIS;
MELITTIN;
GROWTH;
D O I：
10.1016/j.aspen.2017.04.007
中图分类号：
Q96 [昆虫学];
学科分类号：
摘要：
Bee venom has been used for treating various diseases for a long time. However, the bioactive constituents of bee venom and its mechanisms remain poorly understood. In the present study, phospholipase A(2) (Bt-PLA(2)) cDNA was cloned, and a mature form of Bt-PLA2 was purified from bumblebee venom (Bombus terrestris). The differentiation induction and apoptosis induction activities of Bt-PLA(2) on chronic myelogenous leukemia (CML) K562 cells were also evaluated. Bt-PLA(2) cDNA has 540 nucleotides that encode a 180-amino-acid protein. The purified, mature form of Bt-PLA(2) was an 18-kDa protein, and it inhibited K562 cell growth, determined by an IC50 value of 29.5 ng/mu l. Moreover, Bt-PLA(2) induced erythroid differentiation of K562 cells in a dose-dependent manner, and this was supplemented with the upregulation of glycophorin A (GPA) mRNA expression. Bt-PLA(2)-induced apoptosis, analyzed by DAPI staining, was correlated with the result analyzed by AnnexinV-FITC/PI binding. Furthermore, activation of caspase 3 and poly ADP-ribose polymerase (PARP) and inhibition of p-Akt, determined by western blot, further demonstrated that Bt-PLA(2) induced apoptosis mainly through the Akt pathway. The parallel induction of erythroblasts differentiation of K562 cells and apoptosis due to Bt-PLA2 treatment demonstrated the potential use of Bt-PLA(2) as an anti-leukemia drug lead.
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页码：699 / 704
页数：6
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