ADV—miR-184体外转染对人晶状体上皮细胞移行的影响  

ADV-miR-184 Inhibits Human Lens EpitheliaI Cell Migration

作　　者：王晓媛[1] 曹文萍[1] 滕旭[2] 宋武琦[2] 张凤民[2] 刘平[1] 

机构地区：[1]哈尔滨医科大学附属第一临床医学院眼科医院,150001 [2]哈尔滨医科大学微生物教研室,150001

出　　处：《国际遗传学杂志》2009年第5期321-323,332,共4页International Journal of Genetics

基　　金：国家自然科学基金（NSFCJ0730858）

摘　　要：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1．6×10^9puf／mL；分别将ADV—miR-184以MOI10、MOI50、MOI100、MOI200、MOI500转染HLE—B372h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24h、48h、72h、96h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。Objective Investigate the effects of the recombinant adenoviral vector containing microR- NA-184 （ADV-miR-184） on the migration of human lens epithelial cells in vitro. Methods ADV-miR-184 was amplified, purified, and titrated 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 × 10^9puf/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 cells transfected with MOI100 were also significantly inhibited at 24h, 48h and 96h treatments compared with controls （P 〈0. 05）. Conclusion ADV-miR-184 could be successfully used to transfect 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.

关 键 词：人晶状体上皮细胞 miR-184 转染 细胞移行 腺病毒载体 

分 类 号：R776.1[医药卫生—眼科] R973.2[医药卫生—临床医学]