[The effect of acetylcholine and atropine on proliferation and differentiation and mAchR1 expression of human SK-N-SH cells].

Shi yan sheng wu xue bao Pub Date : 2005-08-01
Xin Hu, Jian Xin Lu
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引用次数: 0

Abstract

The SK-N-SH human neuroblastoma cells were treated with acetylcholine (Ach) and atropine (Atro), the proliferation activity of cells was detected by CCK-8 method and the cell cycle distribution was determined by flow cytometry. The changes of the mRNA and protein of mAchR1 and c-fos were analysed by fluorescence quantitative PCR, western blot and flow cytometry indirect-immunofluorescence. Results showed 1 mmol/L Ach could significantly stimulate the proliferation of SK-N-SH cells and 1 mmol/L Atro could prevent SK-N-SH cells from S phase growing into G2/M phase; 1 mmol/L Ach and 1 mmol/L Atro had feedback adjustment on the protein level of mAchR1, but had no significant effect on mRNA level of mAchR1. 1 mmol/L Ach increased the both levels of mRNA and protein of Fos obviously, this effect was reversed by Atro. All phenomena clue us cholinergic receptors joining the events of cell proliferation.

[乙酰胆碱和阿托品对人SK-N-SH细胞增殖分化及mAchR1表达的影响]。
用乙酰胆碱(Ach)和阿托品(Atro)处理SK-N-SH人神经母细胞瘤细胞,CCK-8法检测细胞增殖活性,流式细胞术测定细胞周期分布。采用荧光定量PCR、western blot和流式细胞术间接免疫荧光法分析了mAchR1和c-fos mRNA和蛋白的变化。结果表明,1 mmol/L Ach能显著促进SK-N-SH细胞的增殖,1 mmol/L Atro能阻止SK-N-SH细胞从S期向G2/M期生长;1 mmol/L Ach和1 mmol/L Atro对mAchR1蛋白水平有反馈调节作用,但对mAchR1 mRNA水平无显著影响。1 mmol/L Ach可显著提高Fos mRNA和蛋白水平,但Atro可逆转这一作用。这些现象提示我们胆碱能受体参与了细胞增殖的过程。
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