LncRNA MALAT1在视网膜母细胞瘤组织中的表达及靶向miR-145-5p/SOX9轴对Y79细胞生物行为的影响  

Expression of LncRNA MALAT1 in Retinoblastoma Tissue and Its Effect on Biological Behavior of Y79 Cells by Targeting miR-145-5p/SOX9 Axis

作　　者：严肖啸[1] 尹双慧 李小静 孙敏[4] YAN Xiaoxiao;YIN Shuanghui;LI Xiaojing;SUN Min(Department of Ophthalmology,Handan Central Hospital,Handan 056001,China;Department of Internal Medicine,Handan Hangang Hospital,Handan 056001,China;Department of Orthopedics,the Second Affiliated Hospital of Xingtai Medical College,Xingtai 054002,China;Ophthalmology Department,General Hospital of North China Petroleum Administration Bureau,Cangzhou 062552,China)

机构地区：[1]邯郸市中心医院眼科,邯郸056001 [2]邯郸邯钢医院内科,邯郸056001 [3]邢台医专附属第二医院骨科,邢台054002 [4]华北石油管理局总医院眼科,沧州062552

出　　处：《中国细胞生物学学报》2023年第10期1463-1472,共10页Chinese Journal of Cell Biology

基　　金：河北省2023年度医学科学研究课题计划项目(批准号:20231955)资助的课题。

摘　　要：该文旨在探讨长链非编码RNA(LncRNA)肺腺癌相关转录本1(MALAT1)在视网膜母细胞瘤(RB)组织中的表达情况,及靶向微小RNA(miRNA)-145-5p/性别决定区Y框蛋白9(SOX9)轴对Y79细胞生物行为的影响。qRT-PCR检测RB组织和Y79细胞中MALAT1、miR-145-5p、SOX9mRNA相对表达量;荧光原位杂交(FISH)实验、双荧光素酶报告基因实验验证MALAT1、SOX9与miR-145-5p的靶向关系;将Y79细胞分为sh-NC组、sh-MALAT1组、sh-MALAT1+miR-NC组、sh-MALAT1+miR-145-5p inhibitor组、mimics-NC组、miR-145-5p mimics组、miR-145-5p mimics+pcDNA组、miR-145-5p mimics+SOX9组,MTT法和细胞克隆实验检测细胞增殖;流式细胞术检测细胞凋亡;Transwell小室检测细胞迁移和侵袭;Western blot检测SOX9、Ki67、MMP9、Bcl-2、Cleaved Caspase-3蛋白表达情况。体内肿瘤形成实验验证MALAT1对RB肿瘤的影响。结果显示,MALAT1在RB组织和Y79细胞中高表达(P<0.05);FISH实验、双荧光素酶报告基因实验证实MALAT1、SOX9与miR-145-5p存在靶向关系;沉默MALAT1或过表达miR-145-5p可抑制Y79细胞增殖、迁移和侵袭,促进凋亡(P<0.05);抑制mi R-145-5p或过表达SOX9可逆转沉默MALAT1或过表达miR-145-5p对Y79细胞生物学行为的影响(P<0.05);体内实验显示,抑制MALAT1表达可显著抑制小鼠移植瘤的生长(P<0.05)。MALAT1在RB组织和Y79细胞中高表达,MALAT1可通过调节miR-145-5p/SOX9信号轴,参与Y79细胞生物学行为。The aim of this study was to investigate the expression of LncRNA(long non-coding RNA)MALAT1(metastasis-associated lung adenocarcinoma transcript 1)in RB(retinoblastoma)tissues,and its effect on the biological behavior of Y79 cells by targeting microRNA-145-5p/SOX9(sex determining region Y box protein 9)axis.qRT-PCR was applied to detect the relative expression levels of MALAT1,miR-145-5p,and SOX9 mRNA in RB tissue and Y79 cells.FISH(fluorescence in situ hybridization)experiment and double luciferase reporter gene experiment were applied to verify the targeting relationship between MALAT1,SOX9 and miR-145-5p.Y79 cells were grouped into sh-NC group,sh-MALAT1 group,sh-MALAT1+miR-NC group,sh-MALAT1+miR-145-5p inhibitor group,mimics-NC group,miR-145-5p mimics group,miR-145-5p mimics+pcDNA group,and miR-145-5p mimics+SOX9 group.MTT method and cell cloning experiment were applied to detect cell proliferation.Flow cytometry was applied to detect cell apoptosis.Transwell cells were applied to detect cell migration and invasion.Western blot was applied to detect the expression of SOX9,Ki67,MMP9,Bcl-2,and Cleaved Caspase-3 proteins.In vivo tumor formation experiment was applied to verify the effect of MALAT1 on RB tumors.The results showed that MALAT1 was highly expressed in RB tissue and Y79 cells(P<0.05).FISH experiment and double luciferase reporter gene experiment confirmed that MALAT1,SOX9 and miR-145-5p had a targeting relationship.Silencing MALAT1 or overexpressing miR-145-5p was able to inhibit the proliferation,migration,and invasion of Y79 cells,and promote apoptosis(P<0.05).Inhibiting miR145-5p or overexpressing SOX9 was able to reverse the effects of silencing MALAT1 or overexpressing miR145-5p on the biological behavior of Y79 cells(P<0.05).In vivo experiments showed that inhibiting the expression of MALAT1 was able to obviously inhibit the growth of transplanted tumors in mice(P<0.05).MALAT1 is highly expressed in RB tissue and Y79 cells,and MALAT1 can participate in the biological behavior of Y79 cells by re

关 键 词：视网膜母细胞瘤 生物学行为 LncRNA MALAT1 miR-145-5p 性别决定区Y框蛋白9 

分 类 号：R739.7[医药卫生—肿瘤]