Ifi204基因表达对大鼠血管外膜成纤维细胞凋亡和迁移的影响

Effect of ifi204 gene expression on apoptosis and migration of rat vascular adventitial fibroblast

目的探讨ifi204基因过表达与沉默对大鼠血管外膜成纤维细胞(VAF)凋亡和迁移的影响。方法原代培养大鼠VAF,利用携带ifi204基因的慢病毒颗粒或空载体慢病毒颗粒感染VAF,分别作为实验组(ifi204 lv组)和阴性对照组(blank lv组)。用ifi204基因小干扰RNA瞬时干预VAF使其沉默(ifi204 siRNA组)、空白小干扰RNA作为阴性对照组(control siRNA组),未经处理的VAF作为空白对照组(negative组)。流式细胞仪检测细胞凋亡,细胞划痕法和Transwell法测定细胞迁移情况,用real-time PCR和Western blot分别检测mRNA和蛋白表达。结果与blank lv组、control siRNA组、ifi204 siRNA组和negative组相比,ifi204 lv组的P204、P53 mRNA和蛋白表达及细胞凋亡率明显上调(P<0.05),细胞迁移速度降低(P<0.05)。转染ifi204 siRNA可抑制P204、P53的mRNA和蛋白表达(P<0.05),提高细胞活力,抑制细胞凋亡(P<0.05),加快细胞迁移速度(P<0.05)。结论 ifi204基因表达对大鼠VAF细胞凋亡和迁移的影响可能与激活P53表达有关。

Objective To investigate effects of ifi204 gene over-exprssion and silencing on apoptosis and migration of rat vascular adventitial fibroblast cell（ VAF）. Methods Rat VAFs were cultured and transfected ifi204 gene by lentivirus particles（ ifi204 lv group） or empty vector lentivirus particles（ blank lv group） respectively as the over-exprssion group and blank group. Instantaneous intervention VAF by ifi204 small interfering RNA（ ifi204 siRNA group） or blank small interfering RNA（ control siRNA group） as ifi204 siRNA group and control siRNA group. VAF was untreated as a negative group. Cell apoptosis was detected by flow cytometry; cell migration by cell scratch and transwell; the mRNA and protein expression of P204 / P53 were detected by real-time PCR and Western blot. Results Compared with those in the blank lv group,control siRNA group,ifi204 siRNA group or negative group,in ifi204 lv group,P204 / P53 mRNA and protein expression and rate of apoptosis significantly increased（ P〈0. 05）,cell migration speed reduced（ P〈0. 05）. Transfection of ifi204 siRNA inhibited P204 /P53 mRNA and protein expression（ P〈0. 05）,improved cell viability（ P〈0. 05）,inhibited cell apoptosis（ P〈0. 05）,accelerated the speed of cell migration（ P〈0. 05）. Conclusions Effects of ifi204 gene expression on apoptosis and migration of rat vascular adventitial fibroblast may be related to activation of P53.