Peng LiPan, Chen ZeZhong, Wang GuangChuan, Tian ShuBo, Kong Shuai, Xu Tao, An XiaoHua, Chen YueZhi
Department of Gastrointestinal Surgery, Shandong Provincial Hospital Affiliated to Shandong First Medical University, 250021 Jinan, China.
Department of General Surgery, No. 1 People's Hospital of NingYang County, Taian, 271400 China.
Cancer Cell Int. 2020 Oct 12;20:497. doi: 10.1186/s12935-020-01579-z. eCollection 2020.
Long noncoding RNAs (LncRNAs) have been reported to critically regulate gastric cancer (GC). Recently, it was reported that LBX2 antisense RNA 1 (LBX2-AS1) is abnormally expressed in GC. However, the role of LBX2-AS1 in the malignancy of GC is worth further discussion.
Quantitative real-time polymerase chain reaction (qRT-PCR) was used to determine the LBX2-AS1, miR-4766-5p and C-X-C motif chemokine (CXCL5) expression in GC tissues and cells. Dual-luciferase reporter assay was applied to examine the target relationship between LBX2-AS1 and miR-4766-5p or miR-4766-5p and CXCL5. Cell counting kit-8 (CCK-8) and Transwell assays were used to detect cell proliferation, migration and invasion rates. The protein expression of CXCL5 was confirmed using western blot. The RNA pull down experiment was used to verify the specificity of LBX2-AS1 and miR-4766-5p on BGC-823 and SGC-7901 cells.
LBX2-AS1 was up-regulated in GC tissues and cells, and its knockdown suppressed proliferation, migration and invasion of GC cells. While, overexpression of LBX2-AS1 increased proliferation and increased CXCL5 mRNA level. CXCL5 improved cell proliferation, migration and invasion of GC cells. LBX2-AS1 could bind to miR-4766-5p to regulate CXCL5 expression. Overexpression of CXCL5 overturned those effects of miR-4766-5p in GC cells. RNA Pull down shown that BGC-823 and SGC-7901 cells, miR-4766-5p specifically binds to LBX2-AS1.
In short, this study demonstrated that LBX2-AS1 promoted proliferation, migration and invasion through up-regulation CXCL5 mediated by miR-4766-5p in GC. The LBX2-AS1/miR-4766-5p/CXCL5 regulatory axis provides a theoretical basis for the research on lncRNA-directed therapeutics in GC.
长链非编码RNA(LncRNAs)已被报道在胃癌(GC)中发挥关键调控作用。最近,有报道称LBX2反义RNA 1(LBX2-AS1)在GC中异常表达。然而,LBX2-AS1在GC恶性肿瘤中的作用值得进一步探讨。
采用定量实时聚合酶链反应(qRT-PCR)检测GC组织和细胞中LBX2-AS1、miR-4766-5p和C-X-C基序趋化因子(CXCL5)的表达。应用双荧光素酶报告基因检测法检测LBX2-AS1与miR-4766-5p或miR-4766-5p与CXCL5之间的靶向关系。使用细胞计数试剂盒-8(CCK-8)和Transwell实验检测细胞增殖、迁移和侵袭率。通过蛋白质印迹法确认CXCL5的蛋白表达。采用RNA下拉实验验证LBX2-AS1和miR-4766-5p对BGC-823和SGC-7901细胞的特异性。
LBX2-AS1在GC组织和细胞中上调,其敲低抑制了GC细胞的增殖、迁移和侵袭。同时,LBX2-AS1的过表达增加了细胞增殖并提高了CXCL5 mRNA水平。CXCL5促进了GC细胞的增殖、迁移和侵袭。LBX2-AS1可与miR-4766-5p结合以调节CXCL5表达。CXCL5的过表达逆转了miR-4766-5p在GC细胞中的这些作用。RNA下拉实验表明,在BGC-823和SGC-7901细胞中,miR-4766-5p特异性结合LBX2-AS1。
简而言之,本研究表明LBX2-AS1通过上调miR-4766-5p介导的CXCL5促进GC的增殖、迁移和侵袭。LBX2-AS1/miR-4766-5p/CXCL5调控轴为GC中lncRNA导向治疗的研究提供了理论基础。