摘要
目的:通过建立过表达PC-1的前列腺癌LNCaP细胞系及敲低PC-1表达的C4-2细胞系,探究PC-1激活AKT信号通路的分子机制。方法:将PC-1基因及针对PC-1的siRNA序列,分别克隆至慢病毒表达载体pCDH-EF1-Myc-MCS-T2A-Puro及干扰载体pSIH1-H1-Puro,包装成慢病毒后分别感染前列腺癌LNCaP及C4-2细胞,通过Western印迹鉴定PC-1过表达及敲低效果,并检测PI3K/AKT/mTOR信号通路相关蛋白S6K、AKT的磷酸化水平。结果:PC-1过表达时,S6K磷酸化水平下降,而AKT的磷酸化水平上升。结论:PC-1可以通过抑制S6K激酶活性,解除其对AKT的负反馈抑制作用,从而激活AKT激酶的活性。
Objective： To construct a prostate cancer LNCaP cell line stably overexpressing PC-1 and a C4-2 cell line in which PC-1 expression is stably knocked down and to explore how PC-1 active AKT pathway. Meth-ods： PC-1 and siRNA primers targeting PC-1 were separately cloned into the control vector pCDH-EF1-MCS-T2A-Puro and pSIH1-H1-Puro. Prostate cancer cells LNCaP and C4-2 cells were infected with lentivirus and the expression of PC-1 was identified by Western blot. Meanwhile, Western blot was performed to detect the phosphor-ylation level of PI3K/AKT/mTOR pathway associated proteins such as S6K, AKT. Results： PC-1 overexpression can decrease the S6K phosphorylation level, while increase AKT phophorylation level. Conclusion： PC-1 may ac-tive AKT kinase in some way through releasing negative feedback regulation of S6K on the PI3K/AKT pathway.
出处
《生物技术通讯》
CAS
2014年第4期508-510,共3页
Letters in Biotechnology
基金
国家自然科学基金面上项目(81372740
81372770)
