精准医学杂志2025，Vol.40Issue(4)：290-295,6.DOI:10.13362/j.jpmed.202540076
长链非编码RNA MIATNB对小梁细胞自噬的调控作用及其机制
Regulatory effect of the long non-coding RNA MIATNB on autophagy in trabecular meshwork cells and its mechanism
摘要
Abstract
Objective To investigate the regulatory effect of the long non-coding RNA(lncRNA)MIATNB on autop-hagy in trabecular meshwork cells and its molecular mechanism.Methods RT-qPCR was used to measure the relative expres-sion le-vel of lncRNA MIATNB in iHTM and GTM3 cells.The iHTM cells were divided into A1 group(transfected with siNC)and B1 group(transfected with siMIATNB),and the GTM3 cells were divided into A2 group(transfected with siNC)and B2 group(transfected with siMIATNB);Western blotting(WB)was used to observe the regulatory effect of lncRNA MIATNB on the ex-pression of autophagy-related proteins in the above groups of TM cells.The iHTM cells were divided into C1 group(transfected with siNC)and D1 group(transfected with siMIATNB),and the GTM3 cells were divided into C2 group(transfected with siNC)and D2 group(transfected with siMIATNB);RT-qPCR was used to measure the relative expression level of m6 A-related genes in cells,and RT-qPCR and WB were used to measure the relative mRNA and protein expression levels of IGF2BP1 in iHTM and GTM3 cells.The iHTM cells were divided into E1 group(transfected with siNC)and F1 group(transfected with siIGF2BP1),and the GTM3 cells were divided into E2 group(transfected with siNC)and F2 group(transfected with siIGF2BP1);WB was used to measure the regulatory effect of IGF2BP1 on the expression of LC3B,p62,and Beclin-1 in TM cells,and RNA co-immunoprecipi-tation(RNA-IP)was used to measure the specific binding capacity between the key m6 A reader IGF2BP1 and lncRNA MIATNB.Results Compared with iHTM cells,there was a significant reduc-tion in the relative expression level of lncRNA MIATNB in GTM3 cells(t=4.355,P＜0.05).Compared with group A1/A2,group B1/B2 had a significant increase in LC3B-Ⅱ/Ⅰ ratio(t=3.062,6.399,P＜0.05),a significant increase in the relative expression level of Beclin-1(t=5.454,4.139,P＜0.05),and a significant re-duction in the relative expression level of p62 protein(t=4.799,4.796,P＜0.05).Compared with group C1/C2,group D1/D2 had a significant reduction in the relative expression level of IGF2BP1 in cells(t=3.003,3.832,P＜0.05),and there were no significant differences in the relative expression levels of other m6 A genes between group D1 and group C1(P＞0.05);compared with group C2,group D2 had significant reductions in the expression levels of the METTL14,FTO,ALKBH5,YTHDF1,and YTHDF3 genes(t=2.163-5.294,P＜0.05)and a significant increase in the expression level of the METTL3 gene(t=3.127,P＜0.05).Compared with iHTM cells,GTM3 cells showed significant reductions in the mRNA and protein expression levels of IGF2BP1(t=3.636,3.485,P＜0.05).Compared with group E1/E2,group F1/F2 had a significant increase in LC3B-Ⅱ/Ⅰ ratio(t=10.910,6.399,P＜0.05),a significant increase in the relative expression level of Beclin-1(t=5.533,4.025,P＜0.05),and a signifi-cant reduction in the relative expression level of p62 protein(t=5.035,4.790,P＜0.05).The specific binding capacity between IGF2BP1 and lncRNA MIATNB in GTM3 cells was significantly weaker than that in iHTM cells(t=3.803,P＜0.05).Conclu-sion lncRNA MIATNB inhibits excessive autophagy in TM cells by maintaining the expression of IGF2BP1,and the absent ex-pression of lncRNA MIATNB may be involved in the pathogenesis of glaucoma through m6A.关键词
青光眼/RNA,长链非编码/自噬/RNA结合蛋白质类/小梁细胞/IGF2BP1蛋白,人Key words
Glaucoma/RNA,long noncoding/Autophagy/RNA-binding proteins/Trabecular meshwork cells/IGF2BP1 protein,human分类
医药卫生引用本文复制引用
侯天宇,杨雪娇,张京京,宗瑶,杨淞凯,雷妍蓓,吴若霏,杨先..长链非编码RNA MIATNB对小梁细胞自噬的调控作用及其机制[J].精准医学杂志,2025,40(4):290-295,6.基金项目
青岛市市南区科学技术项目(2022-2-010-YY) （2022-2-010-YY）
青岛大学附属医院临床医学+X科研项目(2023-58) （2023-58）
