MiR-145对视网膜母细胞瘤Y79细胞增殖和凋亡的影响

目的探讨MicroRNA145（miR．145）对视网膜母细胞瘤Y79细胞增殖和凋亡的影响。方法实验研究。将人视网膜母细胞瘤细胞株Y79分为四组：miR-145干预组、阴性miRNA对照组、空脂质体组、空白对照组。脂质体转染法将miR．145转入Y79细胞；荧光定量PCR检测转染后48h成熟miR．145的表达：CCK．8法检测转染48h后的细胞增殖抑制率；流式细胞术检测细胞周期；AnnexinV／PI免疫荧光双染色和流式细胞术检测细胞凋亡。各组数据的比较采用单因素方差分析。结果采用脂质体转染方法．成功将miR．145转染至Y79细胞；荧光定量PCR显示：miR．145干预组成熟miR．145表达（79．06±3．45）明显增加，与阴性miRNA对照组（1．06±0．03）、空脂质体组（O．93±O．02）和空白组（1．00±0．02）比较，差异有统计学意义（F=229．853，P〈0．05）；miR-145干预组的增殖抑制率（21．64％）明显高于阴性miRNA对照组（2．57％）、空脂质体组（3．97％）（F=34．130，P〈0．05）；miR．145抑制Y79细胞周期于G1期：AnnexinV／PI免疫荧光双染色和流式细胞术显示miR．145干预组AnnexinV阳性和AnnexinV／PI双阳性细胞明显增多，阳性率明显高于其他三组，差异有统计学意义（F=35．434，P〈0．05）。结论miR-145可以抑制视网膜母细胞瘤细胞Y79的增殖，诱导细胞凋亡。

Influence of microRNA-145 on the proliferation and apoptosis of retinoblastoma Y79 cells

Objective To observe the expression of MicroRNA 145 （miR-145） in human retinoblastoma Y79 ceils and explore the effect of miR-145 on the proliferation and apoptosis of Y79 cells. Methods In this experimental study, human retinoblastoma Y79 cells were divided into four groups： miR-145 intervention group, control miRNA intervention group, empty liposome group and blank control group, miR-145 was transfected into Y79 cells by lipofection. The expression of miR-145 was detected at 48 hours after transfection by RT-PCR. Cell proliferation inhibition was measured by the Cell Counting Kit-8 （CCK-8）assay. Flow cytometery was used to examine cell cycles. Apoptosis was detected by Annexin/PI double immunofluorescence and flow cytometery. One-way ANOVA was used to analyze the overall data of every group. Results The Y79 cells were transfected by miR-145 successfully with lipofection. RT-PCR analysis showed that the expression of miR-145 in the miR-145 intervention group was significantly increased as compared to the control miRNA intervention group, empty liposome group and blank control group （79.06±3.45 vs 1.06±0.03, 0.93±0.02 and 1.00＋0.02; F=229.853, P〈0.05）. The proliferation inhibition rate was highest in the miR-145 intervention group （21.46%） compared to the control miRNA intervention group （2.57%） and empty liposome group （3.97%） （F=34.130, P〈0.05）. The cell cycle was depressed in the G1 cycle. The results of immunofluorescenee and flow eytometry showed that the ratios of Annexin or Annexin/PI double positive were highest in the miR-145 intervention group, the difference was significant （F=35.434, P〈0.05）. Conclusion miR-145 can inhibit proliferation and induce apoptosis in Y79 cells.