| MicroRNA-4516、MicroRNA-198在视网膜母细胞瘤Y79细胞中的表达及其意义 |
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| 引用本文： | 李萍,卢跃兵,孙爽,刘婷,孙先桃,余继锋.MicroRNA-4516、MicroRNA-198在视网膜母细胞瘤Y79细胞中的表达及其意义[J].眼科新进展,2022,0(3):200-204. |
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| 作者姓名： | 李萍 卢跃兵 孙爽 刘婷 孙先桃 余继锋 |
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| 作者单位： | 450000 河南省郑州市，郑州大学附属儿童医院 河南省儿童医院 郑州儿童医院眼科（李萍，卢跃兵，孙爽，刘婷，孙先桃）；100045 北京市，首都医科大学附属北京儿童医院眼科 国家儿童医学中心（余继锋） |
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| 摘 要： | 目的 探究MicroRNA-4516（miR-4516）、MicroRNA-198（miR-198）在视网膜母细胞瘤（RB）Y79细胞中的表达及其临床意义。方法 收集2018年3月至2021年3月行眼球摘除术治疗的35例RB患儿的肿瘤组织及30例正常视网膜组织标本，比较肿瘤组织、正常视网膜组织和Y79细胞中miR-4516、miR-198的表达水平。脂质体转染法建立miR-4516、miR-198过表达组（miR-4516-mimics组、miR-198-mimics组）、抑制组（miR-4516-inhibitor组、miR-198-inhibitor组）和对照组（miR-4516-NC组、miR-198-NC组）。CCK-8法检测转染24 h、48 h和72 h后各组细胞的增殖活性；实时荧光定量PCR（qRT-PCR）法检测转染48 h后各组细胞miR-4516、miR-198表达水平；流式细胞术检测各组细胞凋亡率；Western blot法检测各组细胞P53、Bcl-2蛋白表达。结果 肿瘤组织及Y79细胞中miR-4516的表达水平均高于正常视网膜组织，miR-198则相反，差异均有统计学意义（均为P<0.05）。细胞转染48 h后，miR-4516、miR-198分别在各自的inhibitor组、NC组和mimics组细胞内相对表达量逐渐升高（均为P<0.05）。转染后24 h、48 h、72 h，各组细胞增殖活性均逐渐升高，差异均有统计学意义（均为P<0.05），且同一时间点各组的细胞增殖活性比较，miR-4516-mimics组>miR-4516-NC组>miR-4516-inhibitor组，miR-198-inhibitor组>miR-198-NC组>miR-198-mimics组，差异均有统计学意义（均为P<0.05）。转染后48 h，各组细胞凋亡率比较，miR-4516-inhibitor组>miR-4516-NC组>miR-4516-mimics组，miR-198-mimics组>miR-198-NC组>miR-198-inhibitor组，差异均有统计学意义（均为P<0.05）。P53蛋白在miR-4516-mimics组、miR-4516-NC组和miR-4516-inhibitor组中表达逐渐升高，Bcl-2蛋白表达逐渐降低，差异均有统计学意义（均为P<0.05）；P53蛋白在miR-198-mimics组、miR-198-NC组、miR-198-inhibitor组中表达逐渐降低，Bcl-2蛋白表达逐渐升高，差异均有统计学意义（均为P<0.05）。结论 miR-4516在RB肿瘤组织及Y79细胞中呈高表达状态，miR-198呈低表达状态；miR-4516可促进Y79细胞增殖，抑制其凋亡，miR-198可抑制Y79细胞增殖，二者有望成为RB的新的肿瘤标志物。
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| 关 键 词： | 视网膜母细胞瘤 MicroRNA-4516 MicroRNA-198 细胞增殖 凋亡 |
Expression of microRNA-4516 and microRNA-198 in retinoblastoma Y79 cells and its significance |
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| LI Ping,LU Yuebing,SUN Shuang,LIU Ting,SUN Xiantao,YU Jifeng.Expression of microRNA-4516 and microRNA-198 in retinoblastoma Y79 cells and its significance[J].Recent Advances in Ophthalmology,2022,0(3):200-204. |
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| Authors: | LI Ping LU Yuebing SUN Shuang LIU Ting SUN Xiantao YU Jifeng |
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| Affiliation: | 1.Department of Ophthalmology,Zhengzhou Children’s Hospital,Henan Children’s Hospital,Zhengzhou University Affiliated Children’s Hospital,Zhengzhou 450000,Henan Province,China2.Department of Ophthalmology,Beijing Children Hospital,Capital Medical University,National Center for Children’s Health,Beijing 100045,China |
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| Abstract: | Objective To investigate the expression of microRNA-4516 (miR-4516) and microRNA-198 (miR-198) in retinoblastoma (RB) Y79 cells and its significance.Methods Thirty-five tumor tissue specimens were collected from children with RB who underwent enucleation of eyeballs between March 2018 and March 2021. Meanwhile, 30 normal retinal tissue specimens were collected. The expression levels of miR-4516 and miR-198 in tumor tissues, normal retinal tissues and Y79 cells were compared. The miR-4516 and miR-198 overexpression groups (miR-4516-mimics group and miR-198-mimics group), inhibitor groups (miR-4516-inhibitor group and miR-198-inhibitor group), and control groups (miR-4516-NC group and miR-198-NC group) were established by lipofection method. The proliferation of cells in each group was detected by CCK-8 assay after 24 h, 48 h and 72 h of transfection. The expression levels of miR-4516 and miR-198 were detected by quantitative real-time polymerase chain reaction after 48 h of transfection. Cell apoptosis was determined by flow cytometry, and the protein expression of P53 and Bcl-2 was measured by Western blot.Results The expression levels of miR-4516 in tumor tissues and Y79 cells were significantly higher than those in normal retinal tissues, while the expression level of miR-198 in tumor tissues and Y79 cells were significantly lower than those in normal retinal tissues(all P<0.05). After 48 h of transfection, the relative expression levels of miR-4516 and miR-198 were significantly increased in the inhibitor groups, NC groups, and mimics groups (all P<0.05). At 24 h,48 h,72 h after transfection, cell proliferation was significantly increased in each group (all P<0.05). The order of cell proliferation at the same time points from high to low was as follows: miR-4516-mimics group > miR-4516-NC group > miR-4516-inhibitor group, miR-198-inhibitor group > miR-198-NC group > miR-198-mimics group (all P<0.05). At 48 h after transfection, the order of cell apoptosis from high to low was as follows: miR-4516-inhibitor group > miR-4516-NC group > miR-4516-mimics group, miR-198-mimics group > miR-198-NC group > miR-198-inhibitor group (all P<0.05). The expression of P53 protein increased, while the expression of Bcl-2 protein decreased in the miR-4516-mimics group, miR-4516-NC group, and miR-4516-inhibitor group (all P<0.05). The expression of P53 protein decreased, while the expression of Bcl-2 protein increased in the miR-198-mimics group, miR-198-NC group, and miR-198-inhibitor group (all P<0.05).Conclusion The expression of miR-4516 is up-regulated and the expression of miR-198 is down-regulated in RB tissues and Y79 cells. MiR-4516 can promote the proliferation and inhibit the apoptosis of Y79 cells, while miR-198 can inhibit the proliferation of Y79 cells. Both are expected to become new tumor markers of RB. |
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| Keywords: | retinoblastoma microRNA-4516 microRNA-198 cell proliferation apoptosis |
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