LncRNA MALAT1调控miR-383-5p对神经母细胞瘤细胞增殖、迁移和侵袭的影响

Effects of LncRNA MALAT1 Regulating miR-383-5p on Proliferation,Migration and Invasion of Neuroblastoma Cells

该文旨在探讨长链非编码RNA(lnc RNA)肺腺癌转移相关转录本1(MALAT1)对神经母细胞瘤(neuroblastoma,NB)细胞增殖、迁移和侵袭的影响及作用机制。选取2019年1月至2021年12月在凉山彝族自治州第一人民医院接受治疗的45例NB患者的NB组织和相邻的非癌组织,利用q RTPCR检测NB组织和相邻的非癌组织、人脐静脉内皮细胞HUVECs和神经母细胞瘤癌细胞系NMB、SHEP21N、SHEP2中MALAT1和miR-383-5p的表达水平。选择SHEP2为研究对象,将其随机分为si-NC组、si-MALAT1组、pc-NC组、pc-MALAT1组、mi R-NC组、mi R-383-5p过表达组(OE-mi R-383-5p组)、anti-NC组、anti-mi R-383-5p组、si-MALAT1+anti-NC组和si-MALAT1+anti-mi R-383-5p组;CCK-8法检测各组SHEP2细胞的增殖水平;Western blot实验检测各组SHEP2细胞中Cyclin D1、PCNA、MMP-2和MMP-9的蛋白表达水平;Transwell实验检测各组SHEP2细胞的迁移和侵袭情况;双荧光素酶报告实验验证MALAT1和mi R-383-5p的靶向关系。NB组织和细胞系(NMB、SHEP21N、SHEP2)中MALAT1表达上调,miR-383-5p表达下调,其中SHEP2细胞中MALAT1表达水平最高,miR-383-5p表达水平最低。敲低MALAT1表达或过表达mi R-383-5p可明显降低SHEP2细胞增殖活性、迁移细胞数目和侵袭细胞数目,并下调Cyclin D1、PCNA、MMP-2和MMP-9蛋白表达水平。MALAT1可靶向调控mi R-383-5p的表达,抑制mi R-383-5p表达可逆转敲低MALAT1对SHEP2细胞增殖、迁移和侵袭的抑制作用。敲低MALAT1可以抑制SHEP2细胞的增殖、迁移和侵袭,其作用机制可能与靶向调控mi R-383-5p有关。

This study is aimed to investigate the effect of lncRNA(long non-coding RNA)MALAT1(metastasis associated in lung denocarcinoma transcript 1)on the proliferation,migration and invasion of NB(neuroblastoma)cells.NB tissues and adjacent non-cancer tissues of 45 NB patients treated in the First People’s Hospital of Liangshan Yi Autonomous Prefecture from January 2019 to December 2021 were selected.The expression levels of MALAT1and miR-383-5p in NB tissue and adjacent non-cancer tissue,human umbilical vein endothelial cells HUVECs and NB cell lines(NMB,SHEP21N,and SHEP2)were detected by qRT-PCR.SHEP2 was seletced as the research object,and was randomly grouped into si-NC group,si-MALAT1 group,pc-NC group,pc-MALAT1 group,miR-NC group,miR-383-5p overexpression group(OE-miR-383-5p group),anti-NC group,anti-miR-383-5p group,si-MALAT1+antiNC group and si-MALAT1+anti-miR-383-5p group.The proliferation of SHEP2 cells in each group was detected by CCK-8 method.The protein expression levels of Cyclin D1,PCNA,MMP-2 and MMP-9 in SHEP2 cells of each group were detected by Western blot assay.The migration and invasion of SHEP2 cells in each group were detected by Transwell assay.Dual luciferase reporter assay verified the targeting relationship between MALAT1 and miR-383-5p.The expression of MALAT1 was up-regulated and miR-383-5p was down-regulated in NB tissues and cell lines(NMB,SHEP21N,SHEP2),with the highest expression of MALAT1 and the lowest expression of miR-383-5p in SHEP2cells.Knockdown of MALAT1 expression or over-expression of miR-383-5p significantly reduced SHEP2 cell proliferation,the number of migrating cells and the number of invasive cells,and down-regulated the expression of Cyclin D1,PCNA,MMP-2 and MMP-9 proteins.MALAT1 can target and regulate the expression of miR-383-5p,and inhibiting the expression of miR-383-5p can reverse the inhibitory effect of knocking down MALAT1 on the proliferation,migration and invasion of SHEP2 cells.The knockdown of MALAT1 can inhibit the proliferation,migration and invasion of SHEP2 cells,and its mechanism of action may be related to the targeted regulation of mi R-383-5p.