lncRNA DSCAM-AS1 通过 miR-144-5p/IRS2轴对甲状腺乳头状癌细胞的影响
Effect of long non-coding RNA DSCAM-AS1 on papillary thyroid carcinoma cells via miR-144-5p/IRS2 axis
吴天思 1陈珊珊 1高博 1石佳宝 1管佳琪 2张小宝 2张立广1
作者信息
- 1. 保定市第二医院 肿瘤外科(河北 保定 071000)
- 2. 承德医学院(河北 承德 067000)
- 折叠
摘要
目的:探究长链非编码RNA(lncRNA)唐氏综合征细胞黏附分子反义1(DSCAM-AS1)通过微小RNA(miR)-144-5p/胰岛素受体底物2(IRS2)轴促进甲状腺乳头状癌(PTC)细胞生长和侵袭的作用.方法:将PTC细胞株TPC-1随机分为对照组(Control组,完全培养基正常培养)、si-NC组(转染si-NC)、si-DSCAM-AS1 组(转染 si-DSCAM-AS1)、si-DSCAM-AS1+inhibitorNC 组(si-DSCAM-AS1 与inhibitor NC 共转染)、si-DSCAM-AS1+miR-144-5p inhibitor 组(si-DSCAM-AS1 与 miR-144-5p inhibitor 共转染).RT-qPCR 法检测 DSCAM-AS1、miR-144-5p 和 IRS2 mRNA 的表达;CCK-8 法检测细胞增殖能力;Transwell实验检测细胞的侵袭能力;Western blot检测IRS2蛋白的表达.双荧光素酶报告基因实验验证靶向关系.结果:与Control组相比,si-DSCAM-AS1组TPC-1细胞DSCAM-AS1表达、吸光度(A450)值、细胞侵袭数目、IRS2表达显著降低(P＜0.05),miR-144-5p表达显著升高(P＜0.05).与 si-DSCAM-AS1 组相比,si-DSCAM-AS1+miR-144-5p inhibitor 组A40值、细胞侵袭数目、IRS2表达显著升高(P＜0.05),miR-144-5p表达显著降低(P＜0.05).DSCAM-AS1靶向负调控miR-144-5p表达,miR-144-5p靶向负调控IRS2表达.结论:沉默DSCAM-AS1可能通过上调miR-144-5p来抑制IRS2蛋白的表达,从而抑制PTC细胞生长和侵袭.
Abstract
Objective:To investigate the role of long non-coding RNA(IncRNA)Down's syn-drome cell adhesion molecule antisense 1(DSCAM-AS1)in promoting the growth and invasion of papillary thyroid carcinoma(PTC)cells through the microRNA(miR)-144-5p/insulin receptor substrate 2(IRS2)axis.Methods:PTC cell line TPC-1 was randomly grouped into Control group(fully cultured in normal medium),si-NC group(transfected with si-NC),si-DSCAM-AS1 group(transfected with si-DSCAM-AS1),si-DSCAM-AS1+inhibitor NC group(co transfected with si-DSCAM-AS1 and in-hibitor NC),and si-DSCAM-AS1+miR-144-5p inhibitor group(co transfected with si-DSCAM-AS1 and miR-144-5p inhibitor).RT-qPCR method was applied to detect the expression of DSCAM-AS1,miR-144-5p,and IRS2 mRNA in cells;CCK-8 method was applied to detect the proliferation ability of cells in each group;Transwell experiment was applied to detect the invasiveness of TPC-1 cells;Western Blot was applied to detect the expression of IRS2 protein.Dual luciferase reporter gene ex-periment was applied to verify of targeting relationship.Results:Compared with the Control group,the expression of DSCAM-AS1,A450 value,number of cell invasions,and the expression of IRS2 in TPC-1 cells in si-DSCAM-AS1 group were obviously reduced(P＜0.05),the expression of miR-144-5p was obviously increased(P＜0.05).Compared with the si-DSCAM-AS1 group,A450 val-ue,number of cell invasions,and the expression of IRS2 in TPC-1 cells in the si-DSCAM-AS1+miR-144-5p inhibitor group were obviously increased(P＜0.05),the expression of miR-144-5p was obviously reduced(P＜0.05).DSCAM-AS1 targeted and negatively regulated miR-144-5p expression,while miR-144-5p targeted and negatively regulated IRS2 expression.Conclusion:Silencing DSCAM-AS1 may inhibit the expression of IRS2 protein by up-regulating miR-144-5p,thereby inhibiting the growth and invasion of PTC cells.
关键词
长链非编码RNA/唐氏综合征细胞黏附分子反义1/miR-144-5p/胰岛素受体底物2/甲状腺乳头状癌/侵袭Key words
Long non-coding RNA Down's syndrome cell adhesion molecule antisense 1/MiR-144-5p/Insulin receptor substrate 2/Thyroid papillary carcinoma/Invasion引用本文复制引用
出版年
2024
NETL
NSTL
万方数据