CAJ | 학술논문

目的探讨MicroRNA 145(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48 h成熟miR-145的表达;CCK-8法检测转染48 h后的细胞增殖抑制率;流式细胞术检测细胞周期;Annexin V/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(0.93±0.02)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;Annexin V/PI免疫荧光双染色和流式细胞术显示miR-145干预组Annexin V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.
목적 탐토MicroRNA 145(miR- 145)대시망막모세포류Y79세포증식화조망적영향.방법 실험연구.장인시망막모세포류세포주Y79분위사조:miR-145간예조、음성miRNA대조조、공지질체조、공백대조조.지질체전염법장miR-145전입Y79세포;형광정량PCR검측전염후48 h성숙miR-145적표체;CCK-8법검측전염48 h후적세포증식억제솔;류식세포술검측세포주기;Annexin V/PI면역형광쌍염색화류식세포술검측세포조망.각조수거적비교채용단인소방차분석.결과 채용지질체전염방법,성공장miR-145전염지Y79세포；형광정량PCR현시:miR-145간예조성숙miR-145표체(79.06±3.45)명현증가,여음성miRNA대조조(1.06±0.03)、공지질체조(0.93±0.02)화공백조(1.00±0.02)비교,차이유통계학의의(F=229.853,P＜0.05); miR-145간예조적증식억제솔(21.64％)명현고우음성miRNA대조조(2.57％)、공지질체조(3.97％)(F=34.130,P＜0.05);miR-145억제Y79세포주기우G1기;Annexin V/PI면역형광쌍염색화류식세포술현시miR-145간예조Annexin V양성화AnnexinV/PI쌍양성세포명현증다,양성솔명현고우기타삼조,차이유통계학의의(F=35.434,P＜0.05).결론 miR-145가이억제시망막모세포류세포Y79적증식,유도세포조망.
Objective To observe the expression of MicroRNA 145 (miR-145) in human retinoblastoma Y79 cells 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 immunofluorescence and flow cytometry 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.