CAJ | 학술논문

目的探讨携带miR-184的重组腺病毒(ADV-miR-184)体外转染对人晶状体上皮细胞移行的影响.方法 ADV-miR-184在293细胞中扩增、纯化并滴定病毒滴度;ADV-miR-184体外感染人晶状体上皮细胞(HLE-B3),采用细胞划痕法测定ADV-miR-184对HLE-B3移行距离的影响.结果测定ADV-miR-184的病毒滴度为1.6×109 puf/mL;分别将ADV-miR-184以MO110、MO150、MOI100、MOI200、MOI500转染HLE-B3 72 h后,细胞移行距离随着ADV-miR-184的浓度增加而减少,MOI100组与MOI50组、MOI10组相比有明显差异(P＜0.05).但与MOI200、MOI500实验组相比变化不明显.与对照组比较,MOI100感染细胞的移行距离在转染后24 h、48 h、72 h、96 h明显缩短(P＜0.05).结论 ADV-miR-184可成功转染人晶体上皮细胞,并可抑制细胞的移行,提示miR可能参与后发性白内障的形成过程.
목적 탐토휴대miR-184적중조선병독(ADV-miR-184)체외전염대인정상체상피세포이행적영향.방법 ADV-miR-184재293세포중확증、순화병적정병독적도;ADV-miR-184체외감염인정상체상피세포(HLE-B3),채용세포화흔법측정ADV-miR-184대HLE-B3이행거리적영향.결과 측정ADV-miR-184적병독적도위1.6×109 puf/mL;분별장ADV-miR-184이MO110、MO150、MOI100、MOI200、MOI500전염HLE-B3 72 h후,세포이행거리수착ADV-miR-184적농도증가이감소,MOI100조여MOI50조、MOI10조상비유명현차이(P＜0.05).단여MOI200、MOI500실험조상비변화불명현.여대조조비교,MOI100감염세포적이행거리재전염후24 h、48 h、72 h、96 h명현축단(P＜0.05).결론 ADV-miR-184가성공전염인정체상피세포,병가억제세포적이행,제시miR가능삼여후발성백내장적형성과정.
Objective Investigate the effects of the recombinant adenoviral vector containing microRNA-184 (ADV-miR-184) on the migration of human lens epithelial cells in vitro.Methods ADV-miR-184 WaS amplified,purified,and titmted in 293 cell line.Human lens epithelial cells (HLE-B3) were then transfected with ADV-miR-184.The) effect of miR-184 on the migration of these transfected cells was evaluated by the scratch assay.Results The original titer of ADV-miR-184 was 1.6×109 puf/mL.HLE-B3 cells were transfected with ADV-miR-184 at different multiplicity of infection (MOI),including MOI10,MOI50,MOI100,MOI200 and MOI500,for a period of 72 h.The migrating distances of HLE-B3 cells were inbitied by MOI100 significantly higher titers of ADV-miR-184 transfection.Transfection with inhibited the migration of HLE-B3 cells compared with groups of control,MOI10,or MOI50 (P＜0.05).In contrast,no differences were showed between MOI100 and MOI200 or MOI500 groups.The migrating distances of HLE-B3 ceils transfected with MOI100 were also significantly inhibited at 24h,48h and 96h tremments compared with controls (P＜0.05).Conclusion ADV-miR-184 could be successfully used to tnmsfect human lens epithelial cells.The miR-184 inhibited the migration of human lens epithelial cells,which suggests that the miR virus may play a role in the development of the posterior capsular opacification.