长链非编码RNA KCNMB2-AS1在胃癌中的表达及机制研究

Expression and mechanism of lncRNA KCNMB2⁃AS1 in gastric cancer

目的探讨长链非编码RNA(long non-coding RNA,lncRNA)KCNMB2-AS1在胃癌(gastric cancer,GC)组织和血浆中的表达水平、临床意义及其对胃癌细胞生物学功能的影响和可能机制。方法实时荧光定量PCR(QRT-PCR)检测82例配对胃癌及癌旁对照组织、31例患者和31例健康体检者血浆KCNMB2-AS1的表达;分析KCNMB2-AS1表达量与临床病理特征的相关性。通过细胞计数、克隆形成、Transwell实验及流式细胞术观察敲减KCNMB2-AS1后胃癌细胞增殖、迁移、侵袭和凋亡的变化。QRT-PCR检测敲减KCNMB2-AS1对肿瘤相关基因表达水平的影响。结果 KCNMB2-AS1在胃癌组织和血浆中显著高表达,其表达水平与肿瘤大小、淋巴结转移、TNM分期及预后密切相关。血浆KCNMB2-AS1表达水平诊断胃癌的ROC曲线下面积为0.865。敲减KCNMB2-AS1抑制胃癌细胞增殖、迁移和侵袭并诱导其凋亡,间质标志N-cadherin、Fibronectin 1和转录因子Twist1、Slug、ZEB1表达下调。结论 KCNMB2-AS1在胃癌组织及血浆中均高表达,其高表达与疾病进展及不良预后相关。血浆KCNMB2-AS1表达水平对诊断胃癌具有一定价值。敲减KCNMB2-AS1抑制胃癌细胞增殖、迁移和侵袭,诱导胃癌细胞凋亡并抑制EMT过程。

Objective To investigate the expression and clinical significance of long non⁃coding RNA(lncRNA)KCNMB2⁃AS1 expression in gastric cancer(GC)tissues and plasma,to explore its effect on the biolog⁃ical functions of GC cells and possible mechanism.Methods 82 paired GC and adjacent control tissues,as well as plasma samples from 34 GC patients and 34 healthy individuals were obtained for real⁃time fluorescence quanti⁃tative polymerase chain reaction(QRT⁃PCR).The correlation between the expression level of KCNMB2⁃AS1 and the clinicopathological features were analyzed.Small interfering RNA(siRNA)was used to knock down KCNMB2⁃AS1 in GC cells.Cell proliferation,migration,invasion and apoptosis assays were then performed to test its biologi⁃cal functions.A series of tumor associated genes in GC cells were detected by QRT⁃PCR after KCNMB2⁃AS1 knock⁃down.Results KCNMB2⁃AS1 was significantly upregulated in GC tissues and plasma.The expression level of KCNMB2⁃AS1 was closely associated with tumor size,lymph node metastasis,TNM stage and prognosis of GC patients.Receiver operating characteristic(ROC)curve analysis showed that the area under the ROC curve(AUC)was 0.865.Knockdown of KCNMB2⁃AS1 inhibited the proliferation,migration and invasion abilities of GC cells and induced their apoptosis.In addition,N⁃cadherin,Fibronectin 1,Twist1,Slug and ZEB1 were downregulated after KCNMB2⁃AS1 knockdown.Conclusions KCNMB2⁃AS1 is upregulated in GC tissues and plasma,which is associated with disease progression and poor prognosis.Plasma expression level showed certain diagnostic value for GC.KCNMB2⁃AS1 knockdown decreased proliferation,migration and invasion of GC cells,while increased apopto⁃sis by inhibiting EMT process.