Objectives: Gastric cancer is a common cancer of the digestive system. Long non-coding RNA (lncRNA) plays an important role in the formation and development of gastric cancer. This study aims to investigate the effect of long non-coding lncRNA 114227 on biologic behaviors in gastric cancer cells.
Methods: The experiment was divided into 4 groups: a negative control (NC) group, a lncRNA 114227 small interference (si-lncRNA 114227) group, an empty vector (Vector) group, and an overexpression vector (OE-lncRNA 114227) group. The expressions of lncRNA 114227 in gastric mucosa and gastric cancer tissues, gastric mucosal epithelial cells and different gastric cancer strains were determined by real-time reverse transcription PCR (real-time RT-PCR).The proliferation were detected by CCK-8 assay in gastric cancer cells. The epithelial-mesenchymal transformation (EMT) was utilized by Transwell assay, scratch healing assay, and Western blotting in gastric cancer cells. The effect of lncRNA 114227 on proliferation of gastric cancer cells was detected by tumor bearing experiment in nude mice in vivo.
Results: The expression level of lncRNA 114227 in the gastric cancer tissues was significantly lower than that in the gastric mucosa tissues, and in 4 kinds of gastric cancer strains was all significantly lower than that in gastric mucosal epithelial cells (all P<0.01). In vitro, the proliferation and migration abilities of gastric cells were significantly reduced after overexpressing lncRNA 114227, and cell proliferation and migration were enhanced after silencing lncRNA 114227 (all P<0.05). The results of in vivo subcutaneous tumorigenesis in nude mice showed that the tumorigenic volume of the tumor-bearing mice in the OE-lncRNA 114227 group was significantly smaller than that of the Vector group, and the tumorigenic quality was lower than that of the Vector group (P<0.05), indicating that lncRNA 114227 inhibited tumorigenesis.
Conclusions: The expression of lncRNA 114227 is downregulated in gastric cancer gastric cancer tissues and cell lines. LncRNA 114227 may inhibit the proliferation and migration of gastric cancer cells through EMT process.
目的: 胃癌是消化系统的常见癌症，患者晚期预后较差。长链非编码RNA(long non-coding RNA，lncRNA)在胃癌的发生、发展中起重要调控作用。本研究主要探讨lncRNA 114227对胃癌细胞生物学行为的影响。方法: 实验分为4组：阴性对照(NC)组、lncRNA 114227小干扰(si-lncRNA 114227)组、空载体(Vector)组、过表达载体(OE-lncRNA 114227)组。采用实时反转录聚合酶链反应(real-time reverse transcription PCR，real-time RT-PCR)检测lncRNA 114227在胃黏膜组织及胃癌组织、胃黏膜上皮细胞及不同胃癌细胞株中的表达情况；采用CCK-8实验检测lncRNA 114227对胃癌细胞增殖的影响；采用Transwell实验、划痕愈合实验和蛋白质印迹法检测lncRNA 114227对胃癌细胞迁移能力和上皮-间充质转化(epithelial-mesenchymal transformation，EMT)相关分子表达的影响；采用裸鼠皮下荷瘤实验检测lncRNA 114227对胃癌细胞体内增殖的影响。结果: LncRNA 114227在胃癌组织的表达水平明显低于胃黏膜组织，在4种胃癌细胞中的表达水平明显低于胃黏膜上皮细胞(均P<0.01)。过表达lncRNA 114227后细胞的增殖和迁移能力明显减弱；沉默lncRNA 114227后细胞的增殖和迁移能力增强(均P<0.05)。体内裸鼠皮下荷瘤实验显示：OE-lncRNA 114227 组过荷瘤小鼠的成瘤体积明显小于Vector 组，成瘤质量低于Vector组(P<0.05)，表明lncRNA 114227抑制肿瘤生成。结论: LncRNA 114227在胃癌组织和胃癌细胞中低表达，可能通过EMT抑制胃癌细胞的增殖和迁移能力。.
Keywords: cell proliferation; epithelial-mesenchymal transformation; gastric cancer; lncRNA 114227.