摘要
目的探讨长链非编码RNA(lncRNA)序列相似家族83成员A-反义核糖核酸1(FAM83AAS1)在视网膜母细胞瘤(RB)细胞中的作用及其潜在的分子机制。方法体外培养RB细胞,将Vector、pcDNA-FAM83A-AS1、NC-siRNA、FAM83A-AS1-siRNA分别转染至细胞中(依次为pcDNA-FAm83A-AS1组、NC-siRNA组、FAm83A-AS1-siRNA组),检测RB细胞中FAM83A-AS1的表达、RB细胞增殖能力、凋亡率。生物信息学方法和双荧光素酶报告基因实验预测和验证FAM83A-AS1与miR-150以及miR-150与高迁移率族蛋白A2(HMGA2)的靶向关系。RT-qPCR检测RB细胞中miR-150、HMGA2 mRNA的表达;Western blot检测RB细胞中HMGA2蛋白的表达;MTT实验和流式细胞术检测转染后RB细胞增殖和凋亡能力。结果pcDNA-FAM83A-AS1组细胞中FAM83A-AS1的表达显著高于Vector组,低于NC-siRNA组(P<0.05)。pcDNA-FAM83A-AS1组较Vector组的细胞增殖能力显著升高,细胞凋亡率显著降低(P<0.05)。FAM83AAS1-siRNA组较NC-siRNA组的细胞增殖能力显著降低,细胞凋亡率显著升高(P<0.05)。miR-150 mimic可显著抑制RB细胞增殖能力,促进细胞凋亡。而pcDNA-HMGA2可显著逆转miR-150 mimic对RB细胞增殖和凋亡的作用。结论敲低lncRNA FAM83A-AS1通过调控miR-150/HMGA2轴抑制RB细胞增殖,促进细胞凋亡。
Objective To investigate the role of long non-coding RNA family with sequence similarity 83 member A antisense RNA 1 in retinoblastoma(RB)cells and its potential molecular mechanism.Methods RB cells were cultured in vitro,and Vector,pcDNA-FAM83A-AS1,NC-siRNA,and FAM83A-AS1-siRNA were transfected into cells to detect the expression of FAM83A-AS1,RB cell proliferation ability,and apoptosis rate in RB cells.Bioinformatics and dual-luciferase reporter gene experiments were performed to predict and verify the targeting relationship between FAM83A-AS1 and miR-150,and miR-150 and the high-mobility family protein A2(HMGA 2).The expression of miR-150 and HMGA2 mRNA in RB cells was measured by RT-qPCR;HMGA 2 protein in RB cells by Western blot;and the proliferation and apoptotic capacity of RB cells after transfection by MTT assay and flow cytometry.Results The expression of FAM83A-AS1 in the pcDNA-FAM83A-AS1 group was significantly higher than that of the Vector group and lower than that of the NC-siRNA group(P<0.05).The cell proliferation capacity in the pcDNA-FAM83A-AS1 group was significantly higher,and the apoptosis rate was significantly lower(P<0.05).Compared with NC-siRNA group,FAM83A-AS1-siRNA group showed significantly increased apoptosis rate(P<0.05).The miR-150 mimic significantly inhibited the proliferation capacity of RB cells and promoted cell apoptosis.However,pcDNA-HMGA 2 significantly reversed the effect of miR-150 mimic on the proliferation and apoptosis of RB cells.Conclusion Knockdown of lncRNA FAM83A-AS1 inhibited RB cell proliferation and promoted apoptosis by regulating the miR-150/HMGA2 axis.
作者
周小平
彭正武
陈书扬
刘茹
田涛
欧玉仑
ZHOU Xiaoping;PENG Zhengwu;CHEN Shuyang;LIU Ru;TIAN Tao;OU Yulun(Department of Ophthalmology,The First People′s Hospital of Chenzhou,Chenzhou 423000,China)
出处
《实用医学杂志》
CAS
北大核心
2023年第22期2934-2939,共6页
The Journal of Practical Medicine
基金
湖南省卫健委科研课题(编号:202207022509)。
