Reismann Marc, Lehmann Daniel, Quandte Matthias, Ure Benno M, Glüer Sylvia
Department of Pediatric Surgery, Hannover Medical School, 30625, Hannover, Germany.
Pediatr Surg Int. 2008 Dec;24(12):1327-30. doi: 10.1007/s00383-008-2274-x.
The aim of the current study was to investigate effects of CO(2) atmosphere, mimicking conditions of the pneumoperitoneum during laparoscopy, on epigenetic conditions of Rassf1A and DCR2 oncogenes in neuroblastoma cells.
SH-SY5Y neuroblastoma cells were exposed to 100% CO(2) for 4 h. Cells were lysed 4, 8 and 168 h after exposure. After methylation analysis of Rassf1A and DCR2 with polymerase chain reaction, results were compared to those of physiologically incubated neuroblastoma cells.
No significant changes were found after exposure to carbon dioxide compared to the control. Values of methylated Rassf1A were 12.6 +/- 1.1 versus 13.2 +/- 1.4 ng/microl in the controls, respectively (4 h after incubation), 12.6 +/- 1.2 versus 15.1 +/- 0.9 ng/microl (8 h) and 14.2 +/- 1.5 versus 11.7 +/- 1.3 ng/microl (168 h). DCR2 showed values of 4.6 +/- 0.5 versus 3.7 +/- 0.5 ng/microl (4 h), 3.8 +/- 0.5 versus 4.1 +/- 0.4 ng/microl (8 h) and 3.6 +/- 0.4 versus 3.8 +/- 0.5 ng/microl (168 h).
Exposure of neuroblastoma cells to 100% CO(2) does not alter methylation of two prognostic relevant index genes. It seems therefore unlikely that effects on methylation levels within CO(2) pneumoperitoneum lead to epigenetic changes in neuroblastoma.
本研究旨在探讨模拟腹腔镜手术中气腹条件的二氧化碳环境对神经母细胞瘤细胞中Rassf1A和DCR2癌基因表观遗传状态的影响。
将SH-SY5Y神经母细胞瘤细胞暴露于100%二氧化碳中4小时。在暴露后4小时、8小时和168小时对细胞进行裂解。在用聚合酶链反应对Rassf1A和DCR2进行甲基化分析后，将结果与生理培养的神经母细胞瘤细胞的结果进行比较。
与对照组相比，暴露于二氧化碳后未发现显著变化。在对照组中，甲基化Rassf1A的值分别为12.6±1.1与13.2±1.4 ng/微升（孵育4小时后）、12.6±1.2与15.1±0.9 ng/微升（8小时）以及14.2±1.5与11.7±1.3 ng/微升（168小时）。DCR2的值分别为4.6±0.5与3.7±0.5 ng/微升（4小时）、3.8±0.5与4.1±0.4 ng/微升（8小时）以及3.6±0.4与3.8±0.5 ng/微升（168小时）。
神经母细胞瘤细胞暴露于100%二氧化碳不会改变两个预后相关指标基因的甲基化。因此，气腹内二氧化碳对甲基化水平的影响似乎不太可能导致神经母细胞瘤的表观遗传变化。
