MicroRNA-34a对人晶状体上皮细胞衰老的影响

Effects of micro RNA-34a on senescence of human lens epithelial cells

目的探讨MicroRNA-34a(MiR-34a)对人晶状体上皮细胞(human lens epithelial cell,HLEC)衰老的影响。方法通过慢病毒介导构建稳定过表达MiR-34a的HLEC-B3细胞株,荧光显微镜下观察细胞转染情况,采用qRT-PCR检测MiR-34的表达。采用β-半乳糖苷酸(SA-β-gal)染色检测HLEC-B3细胞的衰老情况。结果成功转染的细胞显示红色荧光。MiR-34a组MiR-34a的相对表达量(18.69±3.71)显著高于空载体组(1.00±0.02)(P<0.05),成功构建过表达MiR-34a的HLEC-B3细胞株。MiR-34a组和空载体组SA-β-gal染色阳性率分别为(87.67±4.51)%、(7.33±3.21)%,MiR-34a组高于空载体组,差异有统计学意义(P<0.05)。结论过表达MiR-34a促进HLEC衰老,提示MiR-34a可能参与了年龄相关性白内障的形成,有可能成为治疗年龄相关性白内障的新靶点。

Objective To investigate the effects of microRNA-34a（ MiR-34a） on the senescence of human lens epithelial cells（ HLEC）. Methods HLEC-B3 cells were transfected with lentiviral MiR-34 a vector to construct stably over-expressing MiR-34 a cells. The transfection efficiency was confirmed by fluorescence microscope and qRT-PCR. The senescence in HLEC-B3 cells was detected by SA-β-gal staining. Results The transfected cells showed red fluorescence. The expression of MiR-34 a in MiR-34 a group（ 18. 69 ± 3. 71） was higher than that in the vector group（ 1. 00 ± 0. 02）（ P 0. 05）. The stably over-expressing MiR-34 a HLEC-B3 cells was successfully constructed. The SA-β-gal positive rate of MiR-34 a group and vector group were（ 87. 67 ± 4. 51） % and（ 7. 33 ± 3. 21） %,respectively,the SA-β-gal positive rate of MiR-34 a group was significantly higher than that of vector group（ P 0. 05）. Conclusion Over-expression of MiR-34 a in HLEC-B3 cells promote cells senescence,suggesting that MiR-34 a may involve in the formation of cataract and targeting MiR-34 a may be potentially therapeutic approach for the treatment of cataract.