lncRNA CTC-338M12.4抑制miRNA-27a-5p对JAK/STAT信号通路的激活使舌鳞状细胞癌细胞周期阻滞、细胞增殖和迁移受抑制  

Long non-coding RNA CTC-338M12.4 inhibition on activation of JAK/STAT signaling pathway via miRNA-27a-5p makes cell cycle arrest, cell proliferation and migration inhibition in tongue squamous cell carcinoma

作　　者：彭欣 王瑾 熊雁 罗小泉 郭慧 彭建伟 Peng Xin;Wang Jin;Xiong Yan;Luo Xiaoquan;Guo Hui;Peng Jianwei(Department of Stomatology,Third People's Hospital of Hubei Province,Zhongshan Hospital Affiliated to Jianghan University,Wuhan 430071,China;Department of Stomatology,the First People's Hospital of Yunnan Province,Kunming 650034,China)

机构地区：[1]湖北省第三人民医院口腔科,武汉430071 [2]云南省第一人民医院口腔科,昆明650034

出　　处：《肿瘤研究与临床》2024年第11期801-807,共7页Cancer Research and Clinic

基　　金：国家自然科学基金(81860481)。

摘　　要：目的探讨长链非编码RNA(lncRNA)CTC-338M12.4在舌鳞状细胞癌组织中的表达水平及其体外对舌鳞状细胞癌细胞周期、增殖、迁移的作用和分子机制。方法依据基因表达综合(GEO)数据库lncRNA系列数据集GSE139869数据,分析CTC-338M12.4在舌鳞状细胞癌组织和癌旁组织中的差异表达情况。利用反转录实时荧光定量聚合酶链反应(qRT-PCR)检测人永生化口腔角质细胞株HOK及舌鳞状细胞癌细胞株HN13、TSCCa、CAL-27、Tca8113、SCC15中CTC-338M12.4转录水平的表达量。取CTC-338M12.4表达水平最低的CAL-27细胞,分为对照组(转染含阴性序列的载体)和CTC-338M12.4组(转染CTC-338M12.4过表达载体)。采用细胞克隆形成实验检测各组CAL-27细胞增殖能力,采用流式细胞术检测各组CAL-27细胞周期分布,采用划痕实验检测各组CAL-27细胞迁移能力。应用lncACTdb数据库预测CTC-338M12.4与miRNA-27a-5p(miR-27a-5p)有互补结合位点,采用双荧光素酶报告基因实验进行验证。通过qRT-PCR检测各组CAL-27细胞中miR-27a-5p表达水平。通过蛋白质印迹法检测各组CAL-27细胞中JAK/STAT信号通路相关因子的蛋白表达水平。结果对GEO数据库相关数据分析显示,舌鳞状细胞癌组织中转录水平CTC-338M12.4低于癌旁组织,差异有统计学意义(P<0.001)。舌鳞状细胞癌HN13、TSCCa、CAL-27、Tca8113、SCC15细胞中转录水平CTC-338M12.4均低于HOK细胞,差异均有统计学意义(均P<0.05)。CTC-338M12.4组CAL-27细胞中CTC-338M12.4转录水平相对表达量高于对照组,差异有统计学意义(P<0.001)。细胞克隆形成实验显示,CTC-338M12.4组CAL-27细胞克隆数比对照组少[(51±10)个比(114±21)个],差异有统计学意义(t=2.71,P=0.035)。流式细胞术检测显示,CTC-338M12.4组CAL-27细胞中G0/G1期细胞比例高于对照组[(64±3)%比(43±4)%],差异有统计学意义(t=4.87,P=0.003)。划痕实验显示,划痕时两组划痕宽度相近(P>0.05),划痕25 h后CTC-338M12.4组CAL-27�Objective:To investigate the expression level of long non-coding RNA(lncRNA)CTC-338M12.4 in tongue squamous cell carcinoma tissues,and its effects on the cell cycle,cell proliferation,and migration of tongue squamous cell carcinoma cells in vitro as well as its molecular mechanisms.Methods:The Gene Expression Omnibus(GEO)database was used to obtain the lncRNA series data set GSE139869,and the differential expression of CTC-338M12.4 in tongue squamous cell carcinoma tissues and adjacent tissues was analyzed.The transcriptional expression levels of CTC-338M12.4 in human immortalized oral keratinocytes(HOK)and tongue squamous cell carcinoma cell lines HN13,TSCCa,CAL-27,Tca8113,SCC15 were detected by using reverse transcription quantitative real-time polymerase chain reaction(qRT-PCR).CAL-27 cells with the lowest expression level of CTC-338M12.4 were selected and were divided into the control group(co-transfected with vectors containing negative sequence)and CTC-338M12.4 group(co-transfected with CTC-338M12.4 overexpression vectors).The proliferation ability of CAL-27 cells in each group was detected by using cell colony formation assay,and the cell cycle distribution of CAL-27 cells was detected by using flow cytometry.The migration ability of CAL-27 cells was detected by using scratch test.The lncACTdb database was used to predict the complementary binding sites between CTC-338M12.4 and miRNA-27a-5p(miR-27a-5p),and dual-luciferase reporter gene assay was used to verify.The expression level of miR-27a-5p in CAL-27 cells of all groups was detected by using qRT-PCR.The protein expression level of related factors on JAK/STAT signaling pathway in CAL-27 cells of all groups was detected by using Western blot.Results:Analysis of GEO database data showed that transcriptional level CTC-338M12.4 in tongue squamous cell carcinoma tissues was lower than that in adjacent tissues,and the difference was statistically significant(P<0.01).Transcriptional level CTC-338M12.4 in tongue squamous cell carcinoma HNl3,TSCCa,CAL-27,Tca8113

关 键 词：舌肿瘤 癌 鳞状细胞 RNA 长非编码 Janus激酶类 STAT转录因子类 信号传导 CTC-338M12.4 miRNA-27a-5p 

分 类 号：R739.86[医药卫生—肿瘤]