CREST对N2a细胞突起生长的影响

Effect of Calcium-responsive Transactivator on Neurite Outgrowth in N2a Cells

目的观察钙反应性反式激活因子(CREST)过表达对小鼠神经母细胞瘤(N2a)细胞突起生长的影响。方法实验分为3组,实验组用pCMV-HA-CREST转染的N2a细胞,空载体对照组用pCMV-HA转染的N2a细胞,空白对照组用未转染任何试剂的N2a细胞,分别加入70mmol/L KCl孵育过夜。在转染后24h,48h采用考马斯亮蓝染色法检测N2a细胞突起生长状态。结果在转染后24h,CREST基因过表达的细胞与两对照组细胞相比较,细胞突起生长差异无显著性。在转染后48h,CREST基因过表达的细胞与两对照组细胞相比较,突起生长明显,部分突起交织成网络。两对照组间在细胞转染后各时间点突起生长状态差异无显著性(P<0.05)。结论过表达CREST可促进N2a细胞突起的生长。

Objective With the induction of calciumion ,to investigate if overexpression of calcium-respon-sive transactivator（CREST） can promote the growth of neurites in N2a cells.Methods The experiment was carried out in three groups ,and the cells were treated with 70 mmol/L KCl and incubated overnight .In the experimental group ,N2 a cells transfected from CREST gene by pCMV-HA carriers were used;In the non-carrier comparison group , N2a cells transfected from pCMV-HA were used;In the blank control group ,N2a cells which were not transfected were used .The effect of CREST on the outgrowth of neurites of N 2 a cells was assessed by coomassie brilliant blue G-250 colorimetric as-say at 24h and 48h respectively after transfection and observed under phase contrast microscope .Results At 24th hour and 48th hour after the transfection ,CREST was overexpressed in the experimental group compared with the cells in the two comparison groups at two point-in-time;Overexpression of CREST significantly promoted the growth of neurites of N2a cells and the formation of neurite network was also observed at 48th hour after transfection（P＆lt;0.05）.And there was no significant difference between the two control groups .Conclusion Overexpression of calcium-responsive transac-tivator in N2 a cells can promote the growth of neurites .