长链非编码RNARGD1566401在大鼠急性胰腺炎中对外分泌细胞凋亡的作用  

Effect of long non - coding RNA RGD1566401 on apoptosis of rat pancreatic acinar cells in the rat acute pancreatitis model

作　　者：张旭 付强 秦涛 刘传江 胡明星 张宏伟 

机构地区：[1]郑州大学人民医院(河南省人民医院)肝胆胰腺外科,450003

出　　处：《中华实验外科杂志》2018年第1期100-102,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金（31140078、30972928）;河南省科技攻关资助项目（162102310173）

摘　　要：目的观察长链非编码RNA（lncRNA）-RGD1566401（lncR-RGD1566401）在大鼠胰腺腺泡细胞（AR42J）中表达及对AR42J凋亡的影响。方法分别用携带有lncR-RGD1566401目的基因的慢病毒（Lenti-lncRNA）和特异性lncR-RGD1566401抑制剂（lncRNA Smart Silencer）转染AR42J细胞株24 h，转染空病毒（Lenti-NC）和特异性lncR-RGD1566401阴性抑制剂作为对照；以100 nmol/L雨蛙肽（Caerulin）体外诱导正常培养AR42J及转染后AR42J的凋亡，药物诱导24 h后运用流式细胞技术（FCM）检测各组AR42J细胞凋亡率，使用实时荧光定量反转录-聚合酶链反应（RT-qPCR）法检测各组lncR-RGD1566401的表达，采用Western blot检测各组凋亡蛋白活性半胱氨酰天冬氨酸特异性蛋白酶（Caspase）-3和p53表达量变化。结果雨蛙肽体外诱导AR42J凋亡的凋亡率[（19.34±0.54）%]较对照组[（7.23±0.33）%]明显升高，lncR-RGD1566401表达量较对照组升高（5.43±0.31）倍，活性Caspase-3和p53表达量明显升高；转染携带有lncR-RGD1566401目的基因的慢病毒组lncR-RGD1566401表达量较空病毒组升高（8.24±0.22）倍，差异有统计学意义（P＝0.006）；AR42J凋亡率[（38.20±0.37）%]较空病毒组[（22.60±0.96）%]明显升高，差异有统计学意义（P＝0.016）；活性Caspase-3和p53表达量明显升高；应用特异性lncR-RGD1566401抑制剂组较阴性对照组lncR-RGD1566401表达量降低（0.54±0.14）倍，差异有统计学意义（P＝0.003），AR42J凋亡率[（13.60±0.33）%]较阴性对照组[（19.60±0.46）%]明显降低，差异有统计学意义（P＝0.012），活性Caspase-3和p53表达量明显降低。结论lncR-RGD1566401与大鼠AR42J细胞凋亡相关，上调lncR-RGD1566401的表达能够促进大鼠AR42J细胞凋亡。Objective To analyze the expression of RNA-RGD1566401 （long non-coding RNA-RGD1566401, lncR-RGD1566401） in rat pancreatic acinar cells （AR42J） and the effect of lncR-RGD1566401 on apoptosis of AR42J cells.MethodsThe GFP LncR-RGD1566401 overexpressed lentivirus and special long non-coding RNA （lncRNA） Smart Silencer were designed and transfected into AR42J cells repectively for 24 h. The apoptosis of AR42J cells in different groups was established by inducing cells with 100 nmol/L Caerulin for 24 h. The flow cytometry was used to detect the apoptosis of AR42J cells in different groups. Real-time quantitative reverse transcriptase-polymerase chain reaction （RT-qPCR） was used to detect the expression level of lncR-RGD1566401 in AR42J cells of different groups after transfection. The expression of cysteinyl aspartate-specific protease （Caspase）-3 and p53 was detected by Western blotting.ResultsThe apoptosis rate [（19.34±0.54）%] of Caerulin-induced AR42J cells was elevated as compared with the control group [（7.23±0.33）%]. The expression of lncR-RGD1566401 in the AR42J cells was increased by （5.43±0.31） times in the experimental group after treatment with Caerulin and the levels of Caspase-3 and p53 were increased obviously. LncR-RGD1566401 level was elevated by （8.24±0.22） times after AR42J cells were transfected with lenti-lncRNA as compared with the vehicle groups （P=0.006）. The AR42J cells showed more tendency to apoptosis [（38.20±0.37）%] than the vehicle groups [（22.60±0.96）%, P=0.016] and the levels of Caspase-3 and p53 were increased obviously. The AR42J cells that were transfected with Silencer-lncRNA expressed the lower level of lncR-RGD1566401 about [（0.54±0.14）, P=0.003] times and less apoptosis [（13.60±0.33）%] than the Silencer-NC groups [（19.60±0.46）%, P=0.012].ConclusionLncR-RGD1566401 is associated with the apoptosis of AR42J cells, and up-regulation of LncR-RGD1566401 can promote the apoptosis of AR42J cells.

关 键 词：长链非编码RNA 脱噬作用 胰腺腺泡细胞 

分 类 号：R576[医药卫生—消化系统]