长链非编码 RNA ADPGK-AS1 通过激活 ZEB1 介导的上皮-间充质转化促进胰腺癌进展。
LncRNA ADPGK-AS1 promotes pancreatic cancer progression through activating ZEB1-mediated epithelial-mesenchymal transition.
机构信息
a Department of Internal Medicine , Shandong University of Traditional Chinese Medicine , Jinan , Shandong , China.
b Department of Digestive Disease , the Qilu Second Hospital of Shandong University , Jinan , Shandong , China.
出版信息
Cancer Biol Ther. 2018 Jul 3;19(7):573-583. doi: 10.1080/15384047.2018.1423912. Epub 2018 Apr 19.
OBJECTIVE
This study was conducted to investigate the effects of ADP dependent glucokinase antisense RNA 1 (ADPGK-AS1)/ miR-205-5p/ zinc finger E-box binding homeobox 1 (ZEB1) on PC cells.
METHODS
Differentially expressed lncRNAs and miRNAs in pancreatic cancer (PC) were identified by microarray analysis. In silico ceRNA analysis was conducted to find out the interactions among lncRNAs, miRNAs and mRNAs. Quantitative real-time PCR (qRT-PCR) was utilized to examine the expression of miR-205-5p and lncRNA ADPGK-AS1 in PC and non-cancerous cells. The association between miR-205-5p and ADPGK-AS1 as well as miR-205-5p and ZEB1 was determined by dual-luciferase reporter gene assay. After manipulating the expression of ADPGK-AS1, mir-205-5p and ZEB1 in PANC-1 and SW-1990 cells, cell proliferation, migration, invasion and apoptosis were respectively confirmed by cell counting kit-8 (CCK-8) assay, transwell assay and TUNEL. Western blot was applied to examine the expression of Epithelial-mesenchymal Transition-related proteins. In vivo experiment was conducted to further determine the effect of miR-205-5p/ZEB1 on tumorigenic ability of PC cells.
RESULTS
MiR-205-5p was low-expressed while ZEB1 and ADPGK-AS1 were high-expressed in PC tissues and cells compared with the normal. Dual-luciferase reporter gene assay proved that ADPGK-AS1 could directly target miR-205-5p and miR-205-5p could directly target ZEB1 3'UTR. The expression of MiR-205-5p was negatively correlated with proliferation, migration and invasion, and positively correlated with apoptosis rate of PC cells, while ZEB1 and ADPGK-AS1 had an inversed effect. Further in vitro and in vivo investigation indicated that epithelial-mesenchymal transition (EMT) could be restrained by miR-205-5p through targeting ZEB1. ADPGK-AS1 strongly promoted the tumorigenesis via downregulating miR-205-5p expression and induced the EMT process in vivo.
CONCLUSION
ADPGK-AS1 inhibited miR-205-5p and therefore promoted PC progression through activating ZEB1-induced EMT.
目的
本研究旨在探讨 ADP 依赖性葡萄糖激酶反义 RNA 1（ADPGK-AS1）/miR-205-5p/锌指 E 框结合同源盒 1（ZEB1）对 PC 细胞的影响。
方法
通过微阵列分析鉴定胰腺癌（PC）中差异表达的 lncRNA 和 miRNA。通过计算机 ceRNA 分析找出 lncRNA、miRNA 和 mRNA 之间的相互作用。采用实时荧光定量 PCR（qRT-PCR）检测 PC 和非癌细胞中 miR-205-5p 和 lncRNA ADPGK-AS1 的表达。通过双荧光素酶报告基因检测 miR-205-5p 与 ADPGK-AS1 以及 miR-205-5p 与 ZEB1 的相关性。在 PANC-1 和 SW-1990 细胞中转染 ADPGK-AS1、mir-205-5p 和 ZEB1 的表达后，通过细胞计数试剂盒-8（CCK-8）试验、Transwell 试验和 TUNEL 试验分别检测细胞增殖、迁移和侵袭及凋亡情况。采用 Western blot 检测上皮间质转化相关蛋白的表达。通过体内实验进一步确定 miR-205-5p/ZEB1 对 PC 细胞致瘤能力的影响。
结果
与正常组织和细胞相比，PC 组织和细胞中 miR-205-5p 表达降低，ZEB1 和 ADPGK-AS1 表达升高。双荧光素酶报告基因检测证实 ADPGK-AS1 可直接靶向 miR-205-5p，miR-205-5p 可直接靶向 ZEB1 3'UTR。MiR-205-5p 的表达与 PC 细胞的增殖、迁移和侵袭呈负相关，与细胞凋亡率呈正相关，而 ZEB1 和 ADPGK-AS1 的表达则相反。进一步的体外和体内研究表明，miR-205-5p 可通过靶向 ZEB1 抑制上皮间质转化（EMT）。ADPGK-AS1 通过下调 miR-205-5p 的表达强烈促进肿瘤发生，并在体内诱导 EMT 过程。
结论
ADPGK-AS1 通过激活 ZEB1 诱导的 EMT 抑制 miR-205-5p，从而抑制 PC 进展。
Zotero 插件