摘要
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.
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.
出处
《昆明医科大学学报》
CAS
2016年第1期8-11,共4页
Journal of Kunming Medical University
基金
国家自然科学基金资助项目(81260195)
