Inhibition of calmodulin-dependent phosphodiesterase induces apoptosis in human leukemic cells
Inhibition of calmodulin-dependent phosphodiesterase induces apoptosis in human leukemic cells
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DOI：
10.1073/pnas.93.20.11236
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发表时间：
1996-10-01
影响因子：
11.1
通讯作者：
Epstein, PM
中科院分区：
文献类型：
--
作者：
Jiang, X;Li, JP;Epstein, PM
从急性淋巴细胞白血病患者建立的人淋巴母细胞样b细胞系RPMI-8392的细胞质提取物中含有两种主要形式的环核苷酸磷酸二酯酶（PDE）： Ca2+-钙调素依赖性PDE （PDE1）和camp特异性PDE （PDE4）。相比之下，正常的静止人外周血淋巴细胞（HPBL）缺乏PDE1活性[Epstein， P. M., Moraski， S., Jr., and Hachisu， R. (1987) Biochem。[j].科学通报，1999,19(4):444 - 444。利用逆转录聚合酶链反应（RT-PCR），我们发现编码63-kDa形式PDE1 （PDE1B1）的mRNA在rmi -8392细胞中表达，但在正常的静息HPBL中不表达。然而，这种mRNA在植物血凝素（PHA）刺激有丝分裂后在HPBL中被诱导。同样利用RT-PCR技术，从RPMI-8392细胞中克隆了人PDE1B1 cDNA的完整开放阅读框，其编码一个536个氨基酸的蛋白，与牛、大鼠和小鼠的同源性为96%。RT-PCR还鉴定了B- (RPMI-1788, Daudi)和T- （MOLT-4， NA, Jurkat）细胞来源的其他人淋巴母细胞和白血病细胞系中存在PDE1B1，选择性抑制剂抑制PDE1或PDE4活性可诱导RPMI-8392细胞以及其他细胞系发生凋亡。以rpm -8392 mRNA的翻译起始区为靶点，用18 bp的硫代反义寡脱氧核苷酸培养rpm -8392细胞，1天后PDE1B1 mRNA的数量特异性减少，2天后PDE1B1 mRNA的数量消失，并以序列特异性的方式诱导细胞凋亡。这表明PDEs，特别是PDE1B1，由于其表达是选择性的，可能是诱导白血病细胞死亡的有用靶点。
Cytosolic extracts from a human lymphoblastoid B-cell line, RPMI-8392, established from a patient with acute lymphocytic leukemia, contain two major forms of cyclic nucleotide phosphodiesterase (PDE): Ca2+-calmodulin dependent PDE (PDE1) and cAMP-specific PDE (PDE4). In contrast, normal quiescent human peripheral blood lymphocytes (HPBL) are devoid of PDE1 activity [Epstein, P. M., Moraski, S., Jr., and Hachisu, R. (1987) Biochem. J. 243, 533-539]. Using reverse transcription-polymerase chain reaction (RT-PCR), we show that the mRNA encoding the 63-kDa form of PDE1 (PDE1B1) is expressed in RPMI-8392 cells, but not in normal, resting HPBL, This mRNA is, however, induced in HPBL following mitogenic stimulation by phytohemagglutinin (PHA). Also using RT-PCR, the full open reading frame for human PDE1B1 cDNA was cloned from RPMI-8392 cells and it encodes a protein of 536 amino acids with 96% identity to bovine, rat, and mouse species. RT-PCR also identifies the presence of PDE1B1 in other human lymphoblastoid and leukemic cell lines of B- (RPMI-1788, Daudi) and T- (MOLT-4, NA, Jurkat) cell origin, Inhibition of PDE1 or PDE4 activity by selective inhibitors induced RPMI-8392 cells, as well as the other cell lines, to undergo apoptosis. Culture of RPMI-8392 cells with an 18-bp phosphorothioate antisense oligodeoxynucleotide, targeted against the translation initiation region of the RPMI-8392 mRNA, led to a specific reduction in the amount of PDE1B1 mRNA after 1 day, and its disappearance after 2 days, and induced apoptosis in these tells in a sequence specific manner. This suggests that PDEs, particularly PDE1B1, because its expression is selective, may be useful targets for inducing the death of leukemic cells.