HCG22靶向miR-17-3p对口腔鳞癌细胞生物学行为的影响  

Effects of HCG22 on biological behavior of oral squamous cell carcinoma cells by targeting miR-17-3p

作　　者：尹江[1] 唐平[1] 张慧[1] 杨森[1] YIN Jiang;TANG Ping;ZHANG Hui;YANG Sen(Oral Cavity Center,Suining Central Hospital,Suining 629000,China)

机构地区：[1]遂宁市中心医院口腔医学中心,四川遂宁629000

出　　处：《临床肿瘤学杂志》2023年第8期693-699,共7页Chinese Clinical Oncology

摘　　要：目的探讨HLA基因复合体22(HCG22)通过调控微小RNA-17-3p(miR-17-3p)对口腔鳞癌(OSCC)细胞生物学行为的影响。方法通过实时定量PCR(qPCR)检测OSCC组织和细胞系中HCG22和miR-17-3p的表达。双荧光素酶报告基因实验检测HCG22和miR-17-3p之间的相互作用。在OSCC细胞SCC-9中转染pcDNA3.1-HCG22过表达载体和miR-17-3p模拟物mimics,将细胞分为control组(空白)、pcDNA3.1-HCG22组(过表达HCG22)、miR-17-3p mimics组(过表达miR-17-3p)和pcDNA3.1-HCG22+miR-17-3p mimics组(过表达HCG22和miR-17-3p)。MTT、划痕愈合和Transwell法分别检测细胞增殖、迁移和侵袭能力。通过qPCR和Western blotting检测基质金属蛋白酶(MMP)-2和-9的表达。结果OSCC组织中HCG22表达下调,而miR-17-3p表达上调,且两者表达呈负相关(r=-0.794,P<0.05)。与NHOK细胞相比,SCC-15、CAL-27、SCC-6和SCC-9细胞中HCG22低表达,而miR-17-3p高表达(P<0.05)。双荧光素酶报告基因实验证实HCG22能够与miR-17-3p靶向作用。pcDNA3.1-HCG22组SCC-9细胞的miR-17-3p表达水平低于pcDNA3.1组(P<0.05)。与control组比较,pcDNA3.1-HCG22组SCC-9细胞的增殖、迁移和侵袭能力均受到抑制,而miR-17-3p mimics组SCC-9细胞的增殖、迁移和侵袭能力增强。挽救实验显示,miR-17-3p mimics可抵消pcDNA3.1-HCG22诱导SCC-9细胞的增殖、迁移和侵袭的抑制作用。pcDNA3.1-HCG22组的MMP-2和MMP-9水平降低(P<0.05),而miR-17-3p mimics组的MMP-2和MMP-9水平上调(P<0.05)。与pcDNA3.1-HCG22组相比,pcDNA3.1-HCG22+miR-17-3p mimics组的MMP-2和MMP-9水平均升高(P<0.05)。结论HCG22可能通过靶向抑制miR-17-3p表达调控OSCC细胞的增殖、迁移和侵袭过程。HCG22/miR-17-3p轴在OSCC中可能存在负反馈回路。Objective To investigate the function of HLA complex group 22(HCG22)on affecting biological behavior of oral squamous cell carcinoma(OSCC)cells through regulation of microRNA-17-3p(miR-17-3p).Methods The expression levels of HCG22 and miR-17-3p in OSCC tissues and cell lines were determined by qPCR.Dual-luciferase assay was performed to assess the interactions between HCG22 and miR-17-3p.Additionally,pcDNA3.1-HCG22 and miR-17-3p mimics were transfected into SCC-9 cells.Cells were allocated into control(blank),pcDNA3.1-HCG22(overexpression of HCG22),miR-17-3p mimics(overexpression of miR-17-3p)and pcDNA3.1-HCG22+miR-17-3p mimics(overexpression of HCG22 and miR-17-3p)groups.Cell proliferation,migration and invasion were measured by MTT,scratch-healing and Transwell assays.The expression levels of matrix metallopeptidase(MMP)-2 and MMP-9 were detected by qPCR and Western blotting.Results The levels of HCG22 were downregulated and miR-17-3p levels were upregulated in OSCC tissues(P<0.05).Moreover,an inverse correlation between HCG22 and miR-17-3p in OSCC tissues was observed(r=-0.794,P<0.05).Compared with NHOK cells,HCG22 expressions were down-regulated and miR-17-3p expressions were up-regulated in the OSCC cells SCC-15,CAL-27,SCC-6 and SCC-9.HCG22 was predicted to interact with miR-17-3p,which was confirmed by dual-luciferase activity assay.The expression of miR-17-3p in pcDNA3.1-HCG22 group was lower than that in pcDNA3.1 group(P<0.05).The proliferation,migration and invasion abilities of SCC-9 cells in pcDNA3.1-HCG22 group were inhibited compared with control group.While the proliferation,migration and invasion abilities of SCC-9 cells in miR-17-3p mimics group were increased compared with control group.The rescue experiments displayed that miR-17-3p mimics counteracted pcDNA3.1-HCG22 induced inhibition on proliferation,migration and invasion abilities.Levels of MMP-2 and MMP-9 were decreased in pcDNA3.1-HCG22 group(P<0.05)but increased in miR-17-3p mimics group(P<0.05).Levels of MMP-2 and MMP-9 were increased in

关 键 词：口腔鳞癌 HLA基因复合体22 微小RNA-17-3p 迁移 侵袭 

分 类 号：R739.8[医药卫生—肿瘤]