Glyceraldehyde-3-phosphate dehydrogenase antisense oligodeoxynucleotides protect against cytosine arabinonucleoside-induced apoptosis in cultured cerebellar neurons
Glyceraldehyde-3-phosphate dehydrogenase antisense oligodeoxynucleotides protect against cytosine arabinonucleoside-induced apoptosis in cultured cerebellar neurons
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DOI：
10.1073/pnas.93.18.9937
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发表时间：
1996-09-03
影响因子：
11.1
通讯作者：
Chuang, DM
中科院分区：
文献类型：
--
作者：
Ishitani, R;Chuang, DM
阿糖胞苷（AraC）是一种嘧啶抗代谢药，通过抑制DNA合成杀死增殖细胞，重要的是，它也是细胞凋亡的诱导剂。我们最近报道，年龄诱导的培养小脑神经元的凋亡细胞死亡与颗粒状38-kDa蛋白的过度表达直接相关，该蛋白被我们鉴定为甘油醛-3-磷酸脱氢酶（GAPDH; EC 1.2.1.12）。我们发现放线菌素D、放线菌酮或金精三羧酸（一种DNA酶抑制剂）能有效地延缓AraC诱导的未成熟小脑颗粒细胞的神经元死亡。此外，两种GAPDH反义寡脱氧核苷酸能显著抑制AraC诱导的细胞凋亡。这种保护作用比上述经典的凋亡抑制剂更有效。在AraC诱导的神经元死亡之前，GAPDH mRNA水平增加了约2.5倍，并且这种mRNA积累被放线菌素D和GAPDH反义（但非正义）寡核苷酸阻断。与放线菌素D一样，GAPDH反义寡核苷酸也抑制AraC诱导的38-kDa颗粒蛋白的过表达（即，GAPDH），而相应的正义寡核苷酸完全无效。因此，本研究结果表明，GAPDH过表达参与阿糖胞苷诱导的培养小脑颗粒细胞凋亡。
Cytosine arabinonucleoside (AraC) is a pyrimidine antimetabolite that kills proliferating cells by inhibiting DNA synthesis and, importantly, is also an inducer of apoptosis. We recently reported that age-induced apoptotic cell death of cultured cerebellar neurons is directly associated with an over-expression of a particulate 38-kDa protein, identified by us as glyceraldehyde-3-phosphate dehydrogenase (GAPDH; EC 1.2.1.12). We now show that the AraC-induced neuronal death of immature cerebellar granule cells in culture is effectively delayed by actinomycin-D, cycloheximide, or aurintricarboxylic acid (a DNase inhibitor), Furthermore, two GAPDH antisense, but not their corresponding sense, oligodeoxyribonucleotides markedly arrested AraC-induced apoptosis. This protection was more effective than that induced by the above-mentioned classical inhibitors of apoptosis. Prior to AraC-induced neuronal death, GAPDH mRNA levels increased by approximate to 2.5-fold, and this mRNA accumulation was blocked by actinomycin-D and the GAPDH antisense (but not sense) oligonucleotide. Like actinomycin-D, a GAPDH antisense oligonucleotide also suppressed the AraC-induced over-expression of the 38-kDa particulate protein (i.e., GAPDH), while the corresponding sense oligonucleotide was totally ineffective. Thus, the present results show that GAPDH over-expression is involved in AraC-induced apoptosis of cultured cerebellar granule cells.