[1]唐飞,甘露,吕仲平.LINC00473调控miR-210/TET2分子轴对H2O2诱导的人晶状体上皮细胞生物学特性的影响[J].眼科新进展,2020,40(6):533-537.[doi:10.13389/j.cnki.rao.2020.0122]
　TANG Fei,GAN Lu,LYU Zhongping.Effect of LINC00473 on biological characteristics of H2O2 induced human lens epithelial cells by regulating miR-210/TET2 molecularaxis[J].Recent Advances in Ophthalmology,2020,40(6):533-537.[doi:10.13389/j.cnki.rao.2020.0122]
点击复制

LINC00473调控miR-210/TET2分子轴对H2O2诱导的人晶状体上皮细胞生物学特性的影响/HTML
分享到：

《眼科新进展》[ISSN:1003-5141/CN:41-1105/R]

卷:
40卷
期数:
2020年6期
页码:
533-537
栏目:
实验研究
出版日期:
2020-06-05

文章信息/Info

Title:
Effect of LINC00473 on biological characteristics of H2O2 induced human lens epithelial cells by regulating miR-210/TET2 molecularaxis
作者:
唐飞; 甘露; 吕仲平
611731　四川省成都市，四川大学华西医院上锦南府医院眼科（唐飞）； 610000　四川省成都市，华西第四医院眼科（甘露）； 610000　四川省成都市，四川大学华西医院眼科（吕仲平）
Author(s):
TANG Fei1; GAN Lu2; LYU Zhongping3
1.Department of Ophthalmology，Sichuan University Huaxi Hospital Shangjin Nanfu Hospital，Chengdu 611731，Sichuan Province,China
2.Department of Ophthalmology，West China Fourth Hospital，Chengdu 610000，Sichuan Province,China
3.Department of Ophthalmology，West China Hospital of Sichuan University，Chengdu 610000，Sichuan Province,China
关键词:
LINC00473; miR-210; TET2; 过氧化氢; 晶状体上皮细胞; 增殖; 凋亡; 氧化应激
Keywords:
LINC00473;  miR-210;  TET2;  hydrogen peroxide;  lens epithelial cells;  proliferation;  apoptosis;  oxidative stress
分类号:
R776
DOI:
10.13389/j.cnki.rao.2020.0122
文献标志码:
A
摘要:
目的　探讨LINC00473调控miR-210/10-11易位酶2（TET2）分子轴对H2O2诱导的人晶状体上皮细胞生物学特性的影响。方法　人晶状体上皮细胞SRA01/04传代培养后，采用100 μmol·L-1 H2O2处理24 h。将SRA01/04细胞分为Con组、H2O2组、H2O2+pcDNA3.1组、H2O2+pcDNA3.1-LINC00473组、H2O2+pcDNA3.1-LINC00473+miR-NC组、pcDNA3.1-LINC00473+miR-210组、H2O2+pcDNA3.1-LINC00473+si-NC组、H2O2+ pcDNA3.1-LINC00473+si-TET2组。细胞计数试剂盒（CCK-8）检测细胞活力，流式细胞术检测细胞凋亡，试剂盒检测细胞中丙二醛（MDA）含量和超氧化物歧化酶（SOD）、谷光甘肽过氧化物酶（GSH-Px）活性。双荧光素酶报告基因实验检测LINC00473与miR-210、miR-210与TET2的靶向结合关系。结果　Con组与H2O2组、H2O2+pcDNA3.1组与H2O2+pcDNA3.1-LINC00473组、H2O2+pcDNA3.1-LINC00473+miR-NC组与pcDNA3.1-LINC00473+miR-210组、H2O2+pcDNA3.1-LINC00473+si-NC组与H2O2+ pcDNA3.1-LINC00473+si-TET2组相比，细胞存活率、细胞凋亡率、克隆形成数、MDA含量、SOD和GSH-Px活性差异均有统计学意义（均为P<0.05）。双荧光素酶报告基因实验结果显示，miR-NC和WT-LINC00473共转染组SRA01/04细胞荧光素酶活性（0.36±0.03）较miR-210 mimics和WT-LINC00473共转染组（0.96±0.10）显著降低（P<0.05）；miR-NC和WT-TET2共转染组SRA01/04细胞的荧光素酶活性（0.33±0.03）较miR-210 mimics和WT-TET2共转染组（0.94±0.09）显著降低，差异有统计学意义（P<0.05）；LINC00473靶向作用miR-210并下调其表达水平，miR-210可负调控TET2的表达水平。结论　LINC00473通过下调miR-210/TET2分子轴减轻对H2O2对人晶状体上皮细胞增殖、凋亡和氧化损伤的影响。
Abstract:
Objective　To investigate the effect of LINC00473 on the biological characteristics of H2O2 induced human lens epithelial cells by regulating miR-210/10-11 transposase 2 (TET2) molecular axis. Methods　Human lens epithelial SRA01/04 cells were cultured with 100 μmol·L-1 H2O2 for 24 h. SRA01/04 cells were divided into Con group, H2O2 group, H2O2 + pcDNA3.1 group, H2O2 + pcDNA3.1-LINC00473 group, H2O2 + pcDNA3.1-LINC00473 + miR-NC group, pcDNA3.1-LINC00473 + miR-210 group, H2O2 + pcDNA3.1-LINC00473 + si-NC group, and H2O2 + pcDNA3.1-LINC00473 + si-TET2 group. Cell counting kit-8(CCK-8) was used to detect cell viability; flow cytometry was used to detect cell apoptosis; the kit was used to detect malondialdehyde (MDA) content, activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px). The dual luciferase reporter gene assay detected the targeted binding correlation of LINC00473 with miR-210, of miR-210 with TET2. Results　Statistical differences were observed in cell viability, cell apoptosis rate, colony forming efficiency, MDA content, activities of SOD and GSH-Px for Con group comparing with H2O2 group, H2O2 + pcDNA3.1 group comparing with H2O2 + pcDNA3.1-LINC00473 group, H2O2 + pcDNA3.1-LINC00473 + miR-NC group comparing with pcDNA3.1-LINC00473 + miR-210 group, H2O2 + pcDNA3.1-LINC00473 + si-NC group comparing with H2O2 + pcDNA3.1-LINC00473 + si-TET2 group (all P<0.05). The dual luciferase reporter gene confirmed that SRA01/04 cell luciferase activity was 0.36±0.03 in miR-NC and WT-LINC00473 cotransfection group, and obviously lower than 0.96±0.10 in miR-210 mimics and WT-LINC00473 cotransfection group (P<0.05); SRA01/04 cell luciferase activity was 0.33±0.03 in miR-NC and WT-TET2 cotransfection group, and obviously lower than 0.94±0.09 in miR-210 mimics and WT-TET2 cotransfection group (P<0.05). LINC00473 targeted miR-210 and down-regulated its expression level, and miR-210 negatively regulated the expression level of TET2. Conclusion　LINC00473 can reduce the effect of H2O2 on proliferation, apoptosis and oxidative damage of human lens epithelial cells by down-regulating miR-210/ TET2 molecular axis.

参考文献/References:

［1］　SHI Z,SU Y,WANG F,LIU P.Downregulation of microRNA-181a attenuates hydrogen peroxide-induced human lens epithelial cell apoptosis in vitro［J］.Mol Med Rep,2018,17(4):6009-6015.
［2］　FENG K,GUO H K.Eaf2 protects human lens epithelial cells against oxidative stress-induced apoptosis by Wnt signaling［J］.Mol Med Rep,2018,17(2):2795-2802.
［3］　XU X D,LI K R,LI X M,JIN Y,QIN J,YAN B.Long non-coding RNAs:new players in ocular neovascularization［J］.Mol Biol Rep,2014,41(7):4493-505.
［4］　SHEN Y,DONG L F,ZHOU R M,YAO J,SONG Y C,YANG H,et al.Role of long non-coding RNA MIAT in proliferation,apoptosis and migration of lens epithelial cells:a clinical and in vitro study［J］.J Cell Mol Med,2016,20(3):537-548.
［5］　SHI C,YANG Y,YU J,MENG F,ZHANG T,GAO Y.The long noncoding RNA LINC00473,a target of microRNA 34a,promotes tumorigenesis by inhibiting ILF2 degradation in cervical cancer［J］.Am J Cancer Res,2017,7(11):2157-2168.
［6］　ZHOU W Y,ZHANG M M,LIU C,KANG Y,WANG J O,YANG X H.Long noncoding RNA LINC00473 drives the progression of pancreatic cancer via upregulating programmed death-ligand 1 by sponging microRNA-195-5p［J］.J Cell Physiol,2019,234(12):23176-23189.
［7］　WU D,XU Y,ZOU Y,ZUO Q,HUANG S,WANG S,et al.Long noncoding RNA 00473 is involved in preeclampsia by LSD1 binding-regulated TFPI2 transcription in trophoblast cells［J］.Mol Ther Nucleic Acids,2018,12:381-392.
［8］　BAI J,ZHENG Y,DONG L,CAI X,WANG G,LIU P.Inhibition of p38 mitogen-activated protein kinase phosphorylation decreases H2O2-induced apoptosis in human lens epithelial cells［J］.Graefes Arch Clin Exp Ophthalmol,2015,253(11):1933-1940.
［9］　MOK J W,CHANG D J,JOO C K.Antiapoptotic effects of anthocyanin from the seed coat of black soybean against oxidative damage of human lens epithelial cell induced by H2O2［J］.Curr Eye Res,2014,39(11):1090-1098.
［10］　PASCOLINI D,MARIOTTI S P.Global estimates of visual impairment:2010［J］.Br J Ophthalmol,2012,96(5):614-618.
［11］　BAI J,ZHAO W Y,LI W J,YING Z W,JIANG D Q.Long noncoding RNA LINC00473 indicates a poor prognosis of breast cancer and accelerates tumor carcinogenesis by competing endogenous sponging miR-497［J］.Eur Rev Med Pharmacol Sci,2019,23(8):3410-3420.
［12］　ZHANG Q,WANG G,XU L,YAO Z,SONG L.Long non-coding RNA LINC00473 promotes glioma cells proliferation and invasion by impairing miR-637/CDK6 axis［J］.Artif Cells Nanomed Biotechnol,2019,47(1):3896-3903.
［13］　曹佳,严瑾,郜振彦,花静,颜崇淮.姜黄素对铅氧化损伤大鼠的保护作用［J］.中华实用儿科临床杂志,2019,34(12):942-945.
CAO J,YAN J,ZHEN Z Y,HUA J,YAN C H.Protective effect of curcumin on lead oxidative injury in rats ［J］.J Chin Appl Clin Pediat,2019,34(12):942-945.
［14］　王位,马续祥,陈卫东.环黄芪醇对庆大霉素诱导小鼠急性肾损伤的保护作用［J］.新乡医学院学报,2019,36(8):715-719.
WANG W,MA X X,CHEN W D.Protective effect of cycloastragenol on acute kidney injury induced by gentamicin in mice ［J］.J Xinxiang Med Univ,2019,36(8):715-719.
［15］　LU B,CHRISTENSEN I T,MA L W,WANG X L,JIANG L F,WANG C X,et al.miR-24-p53 pathway evoked by oxidative stress promotes lens epithelial cell apoptosis in age-related cataracts［J］.Mol Med Rep,2018,17(4):5021-5028.
［16］　谭钢,吴安花,邵毅,吴恺,邹雪香,周颖.miR-210靶向沉默Bcl-2诱导人晶状体上皮细胞凋亡［J］.眼科新进展,2016,36(8):701-704，708.
TAN G,WU A H,SHAO Y,WU K,ZOU X X,ZHOU Y.miR-210 targeted silence Bcl-2 induces apoptosis of human lens epithelial cells ［J］.Rec Adv Ophthalmol,2016,36(8):701-704,708.

相似文献/References:

[1]谭钢,吴安花,邵毅,等.miR-210靶向沉默Bcl-2诱导人晶状体上皮细胞凋亡[J].眼科新进展,2016,36(8):701.[doi:10.13389/j.cnki.rao.2016.0186]
　TAN Gang,WU An-Hua,SHAO Yi,et al.MiR-210 induced human lens epithelial cell apoptosis by target silencing Bcl-2[J].Recent Advances in Ophthalmology,2016,36(6):701.[doi:10.13389/j.cnki.rao.2016.0186]

备注/Memo

备注/Memo:
N/A
更新日期/Last Update: 2020-06-05