Acetylcholinesterase (AChE) is an important link in the apoptotic pathway induced by hyperglycemia in Y79 retinoblastoma cell line.
Acetylcholinesterase (AChE) is an important link in the apoptotic pathway induced by hyperglycemia in Y79 retinoblastoma cell line.
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DOI：
10.3389/fnmol.2012.00069
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发表时间：
2012
影响因子：
4.8
通讯作者：
Perlman I
中科院分区：
文献类型：
--
作者：
Masha'our RS;Heinrich R;Garzozi HJ;Perlman I
乙酰胆碱酯酶(AChE)在包括视网膜在内的哺乳动物中枢神经系统中被发现在不同类型的应激诱导下表达，从而导致细胞凋亡。在这里，我们测试了AChE在高血糖诱导视网膜细胞系凋亡中的可能参与。Y79视网膜母细胞瘤细胞在饥饿(1%胎牛血清和1 mg/ml葡萄糖)中孵育16~24 h，然后将细胞外葡萄糖浓度提高到3.5 mg/ml或6 mg/ml，以类似水平的甘露醇作为高渗透压的对照。在不同时间段收集细胞，进行细胞凋亡率和AChE蛋白表达分析。免疫印迹法检测PARP的裂解，末端脱氧核苷酸转移酶介导的dUTP缺口末端标记法检测细胞凋亡。用免疫印迹法检测AChE蛋白的表达，用Karnovsky和Roots法检测AChE蛋白的活性。用MissTM乙酰胆碱酯酶shRNA抑制AChE蛋白表达。用3.5 mg/ml葡萄糖处理Y79细胞，而不用3.5 mg/ml甘露醇处理Y79细胞，TUNEL法和PARP裂解实验证实了这一点。部分伴随细胞凋亡的信号通路包括AChE-R变异体和N延伸的AChE变异体的上调，这在mRNA和蛋白水平得到了证实。ShRNA抑制AChE蛋白的表达可保护Y79细胞不进入凋亡途径。我们的数据表明，一个N延伸的AChE变体的表达，很可能是一个R亚型，参与了高血糖引起的Y79细胞的凋亡途径。
Acetylcholinesterase (AChE) expression was found to be induced in the mammalian CNS, including the retina, by different types of stress leading to cellular apoptosis. Here, we tested possible involvement of AChE in hyperglycemia-induced apoptosis in a retinal cell line. Y79 retinoblastoma cells were incubated in starvation media (1% FBS and 1 mg/ml glucose) for 16–24 h, and then exposed to hyperglycemic environment by raising extracellular glucose concentrations to a final level of 3.5 mg/ml or 6 mg/ml. Similar levels of mannitol were used as control for hyperosmolarity. Cells were harvested at different time intervals for analysis of apoptosis and AChE protein expression. Apoptosis was detected by the cleavage of Poly ADP-ribose polymerase (PARP) using western blot, and by Terminal deoxynucleotidyl-transferase-mediated dUTP nick-end-labeling (TUNEL) assay. AChE protein expression and activity was detected by western blot and by the Karnovsky and Roots method, respectively. MissionTM shRNA for AChE was used to inhibit AChE protein expression. Treating Y79 cells with 3.5 mg/ml of glucose, but not with 3.5 mg/ml mannitol, induced apoptosis which was confirmed by TUNEL assay and by cleavage of PARP. A part of the signaling pathway accompanying the apoptotic process involved up-regulation of the AChE-R variant and an N-extended AChE variant as verified at the mRNA and protein level. Inhibition of AChE protein expression by shRNA protected Y79 cell from entering the apoptotic pathway. Our data suggest that expression of an N-extended AChE variant, most probably an R isoform, is involved in the apoptotic pathway caused by hyperglycemia in Y79 cells.
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DOI：
10.1007/bf00304184
发表时间：
1973-01-01
期刊：
HISTOCHEMIE
影响因子：
--
作者：
HANKER, JS;THORNBURG, LP;MOORE, HG
通讯作者：
MOORE, HG
影响因子：
15.9
作者：
Barber, AJ;Lieth, E;Gardner, TW
通讯作者：
Gardner, TW
抗癌药物诱导细胞凋亡过程中 c-Jun-NH2 末端激酶通路介导的乙酰胆碱酯酶表达
影响因子：
8
作者：
Deng, R.;Li, W.;Zhu, X-F
通讯作者：
Zhu, X-F
影响因子：
5.7
作者：
Grisaru, D;Deutsch, V;Soreq, H
通讯作者：
Soreq, H
影响因子：
7.7
作者：
Mohr, S;Xi, X;Kern, TS
通讯作者：
Kern, TS