长链非编码RNA ZEB2-AS1的表达与疾病进展相关，并可预测胃癌患者的预后。

Long non-coding RNA ZEB2-AS1 expression is associated with disease progression and predicts outcome in gastric cancer patients.

作者信息

Xu Chen, Cui Hongli, Li Huazhi, Wu Yongzhe, An Hongchao, Guo Chunhai

机构信息

Department of General Surgery, Chuiyangliu Hospital Affiliated to Tsinghua University, Beijing, China.

出版信息

J BUON. 2019 Mar-Apr;24(2):663-671.

PMID:31128021

Abstract

PURPOSE

To explore the correlation of expression of long non-coding RNA (lncRNA) zinc finger E-box binding homeobox 2 antisense RNA 1 (ZEB2-AS1) in gastric cancer tissues, analyze the correlations of its expression level with the clinicopathological features of gastric cancer and the malignant phenotype of tumor cells, and investigate the molecular mechanism of action of ZEB2-AS1 in gastric cancer.

METHODS

The expression level of ZEB2-AS1 in gastric cancer tissues and cancer-adjacent tissues in 56 patients was detected by fluorescence real-time quantitative polymerase chain reaction (qRT-PCR). AGS human gastric cancer cells were transiently transfected with small interfering lncRNA (si-lncRNA) ZEB2-AS1 using RNA interference technique, and its effect on proliferation of gastric cancer cells was assessed via MTT assay. Hoechst 33342 staining and flow cytometry were performed to examine the effect of ZEB2-AS1 on the apoptosis of AGS cells and scratch and Transwell assay were applied to detect the effect of si-ZEB2-AS1 on the invasion and metastasis of AGS cells.

RESULTS

qRT-PCR results showed that the expression of ZEB2-AS1 in cancer tissues was increased compared with cancer-adjacent tissues. Cell Counting Kit-8 (CCK-8) results indicated that knockdown of ZEB2-AS1 could significantly inhibit the proliferation of AGS cells. Hoechst 33342 staining and flow cytometry demonstrated that knockdown of ZEB2-AS1 obviously promoted the apoptosis of AGS cells. According to scratch and Transwell assay, knocking down ZEB2-AS1 could remarkably inhibit the invasion and metastasis of AGS cells. Western blotting results revealed that knocking down ZEB2-AS1 could inhibit cell invasion and metastasis by suppressing the epithelial to mesenchymal transition (EMT) as well as the expressions of matrix metallopeptidase-2 (MMP-2) and MMP-9 in AGS cells.

CONCLUSIONS

The expression of lncRNA ZEB2-AS1 in gastric cancer tissues is significantly higher than in cancer-adjacent tissues. Patients with highly expressed lncRNA ZEB2-AS1 have a poor prognosis, and knockdown of lncRNA ZEB2-AS1 in AGS cells inhibits cell proliferation, invasion and metastasis, and promotes apoptosis.

摘要

目的

探讨长链非编码RNA（lncRNA）锌指E盒结合同源框2反义RNA1（ZEB2-AS1）在胃癌组织中的表达相关性，分析其表达水平与胃癌临床病理特征及肿瘤细胞恶性表型的关系，并研究ZEB2-AS1在胃癌中的分子作用机制。

方法

采用荧光定量聚合酶链反应（qRT-PCR）检测56例患者胃癌组织及癌旁组织中ZEB2-AS1的表达水平。运用RNA干扰技术将小干扰lncRNA（si-lncRNA）ZEB2-AS1瞬时转染至AGS人胃癌细胞，通过MTT法评估其对胃癌细胞增殖的影响。采用Hoechst 33342染色和流式细胞术检测ZEB2-AS1对AGS细胞凋亡的影响，划痕实验和Transwell实验检测si-ZEB2-AS1对AGS细胞侵袭和转移的影响。

结果

qRT-PCR结果显示，与癌旁组织相比，ZEB2-AS1在癌组织中的表达升高。细胞计数试剂盒-8（CCK-8）结果表明，敲低ZEB2-AS1可显著抑制AGS细胞的增殖。Hoechst 33342染色和流式细胞术表明，敲低ZEB2-AS1明显促进AGS细胞凋亡。根据划痕实验和Transwell实验，敲低ZEB从蛋白免疫印迹结果可知，敲低ZEB2-AS1可通过抑制AGS细胞上皮-间质转化（EMT）以及基质金属蛋白酶-2（MMP-2）和基质金属蛋白酶-9的表达来抑制细胞侵袭和转移。