摘要
为探讨PI3K/Akt信号通路在鸡细胞型朊蛋白(ChPrPC)过表达的DF-1细胞(DF-1-PrP)增殖、黏附、侵袭和凋亡过程中的作用及其与ChPrPC表达量的关系,以DF-1-PrP细胞为模型,空载体转染的DF-1-NC细胞和DF-1细胞为对照,分别用0、10、20、50、100、200nmol·L-1渥曼青霉素处理以抑制PI3K/Akt信号通路,检测细胞对鼠尾胶原的黏附能力,Transwell小室法检测细胞侵袭力,MTT法检测细胞增殖,流式细胞仪检测细胞凋亡,RT-PCR法检测PRNP基因转录量。结果显示,随着渥曼青霉素浓度的增加,DF-1-PrP、DF-1-NC和DF-1细胞的PRNP基因转录量均减少,其增殖、黏附、侵袭能力相应下降,而总凋亡率均升高;但在低于100nmol·L-1的同一渥曼青霉素浓度下,DF-1-PrP细胞增殖、黏附、侵袭能力始终高于对照组,而总凋亡率均低于对照组。本研究表明ChPrPC的过量表达可促进DF-1细胞增殖、黏附和侵袭,抑制其凋亡,在以上过程中,PI3K/Akt信号通路可能具有重要的作用,渥曼青霉素能有效阻断这一通路,但PI3K/Akt信号通路并不是ChPrPC调节细胞凋亡的唯一途径。
DF-1cells with over-expression of chicken PrPC(DF-1-PrP),DF-1cells with pcDNA3.1(DF-1-NC)and DF-1cells were used as cell models in order to study the effect of PI3K/Akt pathway on DF-1-PrP cells proliferation,adhesion,invasion and apoptosis and its relationship with ChPrPC expression.After cells were treated with 0,10,20,50,100 and 200nmol·L-1 Wortmannin,adhesion assay,transwell assay,MTT assay,flow cytometric assay and RT-PCR analyses were used to detect cell adhesion,invasion,proliferation,apoptosis and expression of PRNP mRNA,respectively.The data showed that the expression of PRNP mRNA of DF-1-PrP,DF-1-NC and DF-1cells were all decreased with the increasing of wortmannin concentration,and adhesion,invasion and proliferation ability were reduced,but apoptosis rate were increased.Furthermore,under the same concentration of less than 100nmol·L-1,adhesion,invasion and proliferation ability of DF-1-PrP cells was higher than DF-1-NC and DF-1cells,apoptosis rate was lower.All these results indicated that over-expression of ChPrPC promoted DF-1-PrP cells adhesion,invasion and proliferation and inhibited apoptosis,in which processes PI3K/Akt pathway may play an im-portant role.PI3K/Akt pathway can be effectively blocked by wortmannin,but it may not be the only pathway that ChPrPCregulated cell apoptosis.
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
2015年第10期1844-1850,共7页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
国家自然科学基金(31160510)
甘肃省自然科学研究基金计划项目(1107RJZA198)
甘肃农业大学动物医学院教研产学支持计划(JYCX-KX009)