miR-204对人晶状体上皮细胞移行的影响  

Effect of ADV-miR-204 on human lens epithelial cell migration

作　　者：王晓媛[1] 白洁[1] 丛丽丹[1] 高维奇[1] 宋武琦[2] 张凤民[2] 

机构地区：[1]哈尔滨医科大学附属第一医院眼科 [2]哈尔滨医科大学微生物学教研室

出　　处：《哈尔滨医科大学学报》2017年第5期413-416,共4页Journal of Harbin Medical University

基　　金：黑龙江省教育厅科学技术研究项目(12521301)

摘　　要：目的探讨经体外转染携带有miR-204的重组腺病毒ADV-miR-204对人晶状体上皮细胞移行的影响。方法 ADV-miR-204在293细胞中扩增、分离纯化并滴定病毒滴度;ADV-miR-204体外转染人晶体上皮细胞HLE-B3,细胞划痕法测定ADV-miR-204对HLE-B3移行的影响。结果测定ADVmiR-204的病毒滴度为1.5×109pfu/m L;ADV-miR-204以MOI为10、50、100、200、500分别感染HLE-B372 h后,细胞移行愈合率随着ADV-miR-204的感染浓度增加而降低,MOI=100组与MOI=50组、MOI=10组相比差异显著(P<0.05)。而在MOI=100、MOI=200、MOI=500实验组组间差异不明显。同一感染浓度(MOI=100),不同时间点(0~72 h)下感染组细胞移行愈合率与正常细胞组比较均明显降低(P<0.05)。结论 ADV-miR-204可成功转染人晶体上皮细胞,并可减缓细胞的移行。Objective To investigate the effects of the recombinant adenoviral vector contai- ning microRNA-204 （ADV-miR-204） on the migration of human lens epithelial cells in vitro. Methods ADV-miR-204 was amplified, purified, and titrated in 293 cell line. Human lens epithelial cells （HLE-B3） were infected by ADV-miR-204. The migration distance of infected HLE-B3 was determined by scratch assay. Results The original titer of ADV-miR-204 was 1.5 x 109 pfu/mL. After HLE-B3 cells were transfected with ADV-miR-204 in different multi- plicity of infection （MOI）, including MOI = 10, MOI = 50, MOI = 100, MOI = 200 and MOI = 500, for a period of 72 h, the ratio of migrated heal of HLE-B3 shortened with increasing ADV-miR-204＇s concentration. The ratio of migrated heal of HLE-B3 cells in transfection group in MOI = 100 was significantly decreased compared with that in MOI = 10 and MOI = 50 respectively （P 〈 0.05 ）. In contrast, no differences were showed among the groups of MOI = 100, MOI = 200 and MOI = 500. The ratio of migrated heal of transfection groups in MOI = 100 was apparently decreased compared with the control groups at different time points respectively （P 〈 0. 05 ）. Conclusion ADV-miR-204 could be successfully used to transfect HLE-B3.The miR-204 could inhibit the migration of HLE-B3.

关 键 词：人晶体上皮细胞 腺病毒 miR-204 转染 细胞移行 

分 类 号：R770.5[医药卫生—眼科]