TMED2受miR-5583-5p调控在舌鳞状细胞癌细胞中发挥癌基因作用

TMED2 is Regulated by miR-5583-5p to Elicit an Oncogene Effect in Tongue Squamous Cell Carcinoma

舌鳞状细胞癌(tongue squamous cell carcinoma,TSCC)是口腔鳞状细胞癌(oral squamous cell carcinoma,OSCC)中最常见类型,其发病率和死亡率都较高。大量研究表明,microRNA(miRNA)是一类小非编码RNA,可调节靶基因的转录后处理,导致靶mRNA的降解及翻译抑制。然而,关于跨膜p24转运蛋白2(transmembrane p24 trafficking protein 2,TMED2)受miR-5583-5p调控对TSCC细胞Cal-27迁移、侵袭、增殖及上皮间充质转化(epithelial-mesenchymal transition,EMT)能力影响尚不明确。本研究利用数据库,分析TMED2高表达头颈部肿瘤(head and neck cancer,HNC)病人愈后不良,在HNSCC组织中的表达升高(P<0.001)。Western印迹结果表明,在6例TSCC组织及TSCC细胞系SCC-9、SCC-25和Cal-27中,TMED2蛋白质表达量较癌旁组织及口腔正常上皮细胞中上调,转染TMED2干扰质粒(SiTMED2)的Cal-27细胞中TMED2蛋白质表达量降低,E-钙黏着蛋白(E-cadherin)表达上调,而N-钙黏着蛋白(N-cadherin)和波形蛋白(vimentin)表达下调。迁移与侵袭结果发现,转染SiTMED2的Cal-27细胞穿过小室基底膜细胞数量均低于对照组(P<0.05)。EdU结果显示,转染SiTMED2的Cal-27细胞增殖能力降低(P<0.05)。双荧光素酶结果显示,TMED2存在与miR-5583-5p的结合靶点。荧光定量RT-PCR结果显示,miR-5583-5p在Cal-27细胞中表达量较Hoec细胞中下降,转染过表达miR-5583-5p质粒的Cal-27细胞其表达量升高(P<0.05),过表达miR-5583-5p(miR-5583-5p mimics)后TMED2蛋白表达水平降低(P<0.05)。综上所述,TMED2受miR-5583-5p调控促进舌鳞状细胞癌细胞Cal-27迁移、侵袭、增殖及上皮间充质转化的发生。

Tongue squamous cell carcinoma(TSCC)is the most common type of oral squamous cell carcinoma(OSCC)with high morbidity and mortality.Many studies have shown that microRNA(miRNA)are small non-coding RNA that regulate the post-transcriptional processing of target genes,resulting in the degradation and translation inhibition of target mRNA.However,how the transmembrane p24 trafficking protein 2(TMED2)is regulated by miR-5583-5 p on migration,invasion,proliferation and epithelial-mesenchymal transition(EMT)of TSCC Cal-27 cells is unclear.In this study,a database was used to analyze the expression of TMED2 in HNSCC(P<0.001)in head and neck cancer(HNC).Western blot showed that the expression of TMED2 protein was up-regulated in 6 cases of TSCC tissues and cell lines such as SCC-9,SCC-25 and CAL-27.After the Cal-27 cells transfected with TMED2 interference plasmid(SiTMED2)the expression of E-cadherin was up-regulated,and N-cadherin and Vimentin was down-regulated.Migration and invasion experiments showed that the number of cells transfused into the basement membrane of the cells was lower than that of the control group(P<0.05).The results of EdU showed that the proliferation of Cal-27 cells transfected with SiTMED2 was decreased(P<0.05).The results of dual luciferase experiment showed that TMED2 had a binding target to miR-5583-5 p,and the expression of miR-5583-5 p in Cal-27 cell was lower than that in Hoec cells.The expression of miR-5583-5 p was increased and TMED2 protein was decreased after the Cal-27 cells were transfected with miR-5583-5 p plasmid(P<0.05).In conclusion,TMED2 is regulated by miR-5583-5 p and promoted the migration,invasion,proliferation and EMT of tongue squamous cell carcinoma cell Cal-27.