Mechanism of long non-coding RNA-metastasis associated lung adenocarcinoma transcript 1 induced invasion and metastasis of esophageal cancer cell EC-109
10.3760/cma.j.issn.0253-3766.2017.06.002
- VernacularTitle: 长链非编码RNA－肺腺癌转移相关转录子1调控食管癌EC－109细胞侵袭转移的机制
- Author:
Qingqin ZHANG
1
;
Yanhui CUI
1
;
Ying WANG
1
;
Weizheng KOU
1
;
Fei CAO
2
;
Xiangjun CAO
2
;
Zhanhui MIAO
1
;
Xiaohong KANG
1
Author Information
1. Department of Oncology, the First Affiliated Hospital of Xinxiang Medical University, Weihui 453100, China
2. Department of Orthopaedics, Pingdingshan First People′s Hospital, Pingdingshan 467000, China
- Publication Type:Journal Article
- Keywords:
Esophageal neoplasms;
Long non-coding RNA;
MicroRNA;
MALAT1;
Epithelial-mesenchymal transition, EMT;
Invasion
- From:
Chinese Journal of Oncology
2017;39(6):405-411
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the effect and mechanism of long non-coding RNA-metastasis associated lung adenocarcinoma transcript 1, (LncRNA-MALAT1) on invasion and metastasis of esophageal cancer cell EC-109.
Methods:EC-109 cells were transfected with lentiviral vector carrying short hairpin RNA of MALAT1( shRNA-MALAT1) or a nonspecific shRNA control (shRNA-control). The expressions of MALAT1, microRNA-200a, ZEB1 and ZEB2 were detected by qRT-PCR. The effect of shRNA-MALAT1 on invasion of EC-109 cells was determined by transwell assay. The expressions of components of epithelial-msenchymal transition pathway in EC-109 cells were determined by immunofluorescence array and western blotting. The expression relationship between MALAT1 and miR-200a in EC-109 cells was detected by dual-luciferase reporter assay.
Results:The result of qRT-PCR showed that the expressions levels of MALAT1, ZEB1 and ZEB2 in shRNA-MALAT1 group were 0.43±0.06, 0.64±0.04 and 0.51±0.04, respectively, significantly lower than 0.97±0.08, 1.06±0.07 and 0.98±0.05 in shRNA-control group and 1 in control group, respectively(all P<0.05). Transwell assay showed that the number of invaded cells in shRNA MALAT1 group was (96.81±10.43) per low-power field, markedly lower than that of (278.44±13.28) per low-power field in shRNA-control group (P<0.01). Immunofluorescence staining and Western blotting showed that MALAT1 downregulation significantly reduced the expressions of proteins related to EMT signal pathway in EC-109 cells.Dual luciferase reporter assay showed that compared to negative control, the activities of luciferase reporter in EC-109 cells co-transfected with pmirGLO-MALAT1-wt and miR-200a were significantly down-regulated. While co-transfected pmirGLO-MALAT1-mut with miR-200a mimics had no effect on the luciferase reporter activities of MALAT1.
Conclusion:LncRNA MALAT1 functions as a competing endogenous RNA to regulate the expressions of ZEB1 and ZEB2 by sponging miR-200a and promotes invasion and migration of esophageal cancer cells through inducing epithelial-mesenchymal transition.
