Gnaq对SH-SY5Y细胞增殖的作用及机制

目的研究G蛋白α亚基Gnaq对人神经母细胞瘤细胞株SH-SY5Y增殖的影响及其可能机制.方法利用慢病毒转染技术在SH-SY5Y细胞中过表达Gnaq,利用荧光显微镜检测转染效率,利用逆转录PCR检测过表达后Gnaq的转录水平,利用MTT技术检测过表达Gnaq后细胞增殖速率的变化,同时利用免疫印迹实验检测转录因子NF-κB的表达水平的变化,分析Gnaq影响SH-SY5Y细胞增殖速率的可能机制.结果 （1）慢病毒载体对SH-SY5Y细胞的转染效率达（97.60±2.14）%;（2）过表达Gnaq基因后,SH-SY5Y细胞中的Gnaq转录水平明显升高,细胞的增殖速率明显加快,NF-κB表达水平明显上调...

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Published in昆明医科大学学报 Vol. 37; no. 1; pp. 8 - 11
Main Author 贾楠楠 邹智荣 李国萍 叶频 刘承杏 陈绍春
Format Journal Article
LanguageChinese
Published 2016
Subjects
Gnaq
NF-κB
SH-SY5Y细胞
增殖
机制
Online AccessGet full text
ISSN2095-610X

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Summary:目的研究G蛋白α亚基Gnaq对人神经母细胞瘤细胞株SH-SY5Y增殖的影响及其可能机制.方法利用慢病毒转染技术在SH-SY5Y细胞中过表达Gnaq,利用荧光显微镜检测转染效率,利用逆转录PCR检测过表达后Gnaq的转录水平,利用MTT技术检测过表达Gnaq后细胞增殖速率的变化,同时利用免疫印迹实验检测转录因子NF-κB的表达水平的变化,分析Gnaq影响SH-SY5Y细胞增殖速率的可能机制.结果 （1）慢病毒载体对SH-SY5Y细胞的转染效率达（97.60±2.14）%;（2）过表达Gnaq基因后,SH-SY5Y细胞中的Gnaq转录水平明显升高,细胞的增殖速率明显加快,NF-κB表达水平明显上调（P〈0.05）.结论 Gnaq基因通过调节NF-κB信号通路而对SH-SY5Y细胞发挥明显的促增殖作用.
Bibliography:Objective To study the role of Gnaq, the α subunit of G proteins, in the proliferation of neuroblastoma cell line SH-SY5Y,and further explore the underlying mechanism. Methods Over-expression of Gnaq in SH-SY5Y cells was performed by lentivirus infection. The infection rate was checked by fluorescence microscope. Then MTT experiment was used to evaluate the proliferation of the Gnaq-overexpressed cells. The transcription level of Gnaq and expression level of nuclear factor kappa B（NF-κB） were measured by PCR and Western Blot respectively. Results（1） The infection rate of lentivirus in SH-SY5Y cells was（97.60±2.14） %.（2） The proliferation and NF-κB level of SH-SY5Y cells was significantly upregulated with Gnaq over-expression.Conclusions Gnaq plays an important role in the proliferation of SH-SY5Y cells by regulating the NF-κB signal pathway.
53-1221/R
JIA Nan-nan;ZOU Zhi-rong;LI Guo-ping;YE Pin;LIU Cheng-xing;CHEN Shao-chun; （1） Dept. of Anatomy and Histoembryology, Kunming Medical University, Kunming Yunnan 65
ISSN:2095-610X