摘要
目的观察TIPE2稳定沉默后对TLR激动剂Poly I:C诱导THP-1凋亡的影响。方法以THP-1细胞为研究对象,使用不同TLR激活剂与THP-1细胞孵育后,采用荧光定量PCR检测TIPE2 m RNA表达。应用慢病毒技术稳定沉默TIPE2表达后,MTT法检测细胞增殖,Annexin V/PI双染法检测细胞凋亡,免疫印迹分析TIPE2沉默后Caspase-8表达水平变化。结果 TLRs激活剂中TLR3特异性激活剂Poly I:C可以显著上调TIPE2的表达(P<0.05),随作用时间延长,可显著抑制THP-1细胞增殖,并可诱导细胞凋亡。利用慢病毒技术稳定沉默THP-1细胞中TIPE2表达以后,可显著削弱Poly I:C诱导的TIPE2表达(P<0.05),TIPE2沉默后,Poly I:C抑制THP-1增殖和诱导细胞凋亡的效应显著削弱,随后免疫印迹的结果显示,Caspase-8的活化形式,p41/42表达下调。结论Poly I:C通过上调TIPE2表达可抑制THP-1细胞增殖,并可诱导细胞凋亡,稳定沉默TIPE2后可一定程度削弱上述效应,可能与下调Caspase-8的活性形式表达有关。
Objective To investigate the influence of stably silent TIPE2 on THP-1 apoptosis induced by TLR agonist Poly I：C. Methods THP-1 cells were selected as the study subjects. The expression of TIPE2 mRNA was detected by fluorogenic quantitative PCR after incubation of THP-1 cells with different TLR activating agents. After stable silence of TIPE2 by lentiviral technique, the cell proliferation was detected by MTI＇, apoptosis was detected by Annexin V/PI double-staining method, and changes of Caspase-8 expression level after silence of TIPE2 was analyzed by western blot. Results TLR3 specific activator Poly I：C could significantly up-regulate the expression of TIPE2（P〈0.05）. Along with the time of action, it could significantly inhibit the proliferation of THP-1, and induce apoptosis. Stable silence of TIPE2 in THP-1 by lentiviral technique could significantly impair the TIPE2 expression induced by Poly I!C （P〈0.05）. After silence of TIPE2, the effects of Poly I：C on inhibition the proliferation of THP-1 and induction of apoptosis were significantly impaired, and the result of following western blot showed active form of Caspase-8 and down-regulation of p41/42 expression. Conclusion By up-regulating the expression of TIPE2, Poly I：C can inhibit the proliferation of THP-1, and induce apoptosis, which can be partially impaired by stable silence of TIPE2, possibly due to down-regu- lation of Caspase-8 active form expression.
出处
《中国现代医生》
2016年第16期1-5,F0003,共6页
China Modern Doctor
基金
浙江省宁波市社会发展基金支持(2013C50043)
