Silencing lncRNA AFAP1-AS1 Inhibits the Progression of Esophageal Squamous Cell Carcinoma Cells via Regulating the miR-498/VEGFA Axis
Authors Shen W, Yu L, Cong A, Yang S, Wang P, Han G, Gu B, Zhang W
Received 17 March 2020
Accepted for publication 11 July 2020
Published 29 July 2020 Volume 2020:12 Pages 6397—6409
Checked for plagiarism Yes
Review by Single-blind
Peer reviewer comments 4
Editor who approved publication: Dr Kenan Onel
Wenhao Shen,1,2,* Lei Yu,1,2,* Aihua Cong,1,2 Song Yang,1,2 Peng Wang,1,2 Gaohua Han,1,2 Bin Gu,2,3 Wei Zhang2,4
1Department of Oncology, Taizhou People’s Hospital, Taizhou, Jiangsu, People’s Republic of China; 2Medical School of Nantong University, Nantong, Jiangsu, People’s Republic of China; 3Department of Emergency, Taizhou People’s Hospital, Taizhou, Jiangsu, People’s Republic of China; 4Department of Infectious Disease, Taizhou People’s Hospital, Taizhou, Jiangsu, People’s Republic of China
*These authors contributed equally to this work
Correspondence: Wei Zhang Department of Infectious Disease, No. 399, South Hailing Road, Taizhou 225300, Jiangsu, People’s Republic of China
Bin Gu Department of Emergency, No. 399, South Hailing Road, Taizhou 225300, Jiangsu, People’s Republic of China
Purpose: In view of the continuous increase of the mortality rate, esophageal squamous cell carcinoma (ESCC) develops into a major health concern. In this study, we aimed to investigate the underlying mechanism of long noncoding RNA (lncRNA) actin filament-associated protein 1 antisense RNA (AFAP1-AS1)/microRNA-498 (miR-498)/vascular endothelial growth factor A (VEGFA) in ESCC cells.
Methods: The expression levels of AFAP1-AS1, miR-498 and VEGFA in ESCC tissues and cells were detected using quantitative real-time polymerase chain reaction (qRT-PCR). The effects of AFAP1-AS1 on ESCC cells proliferation and apoptosis were measured by methyl thiazolyl tetrazolium (MTT) and flow cytometry, respectively. Transwell assay was carried out to determine cell migration. In addition, VEGFA and cell behaviors-related proteins were determined by Western blot analysis. The targeted relationships of AFAP1-AS1 were verified by dual-luciferase reporter and RNA pull-down assays.
Results: The expression levels of lncRNA AFAP1-AS1 and VEGFA mRNA were upregulated, but miR-498 was downregulated in ESCC tissues and cells. Moreover, miR-498 was directly targeted by AFAP1-AS1 and there was a negative correlation between miR-498 and AFAP1-AS1. Functionally, AFAP1-AS1 silencing inhibited the proliferation and migration and induced apoptosis of ESCC cells. Interestingly, miR-498 inhibition rescued the effects of AFAP1-AS1 knockdown on cell proliferation, apoptosis and migration and restored the expression levels of tumor-developing marker proteins of AFAP1-AS1 silencing in Eca109 and KYSE-30 cells. Furthermore, VEGFA was verified as a direct target of miR-498 and reversed the effects of miR-498 overexpression on cell behaviors of ESCC in vitro.
Conclusion: Downregulation of AFAP1-AS1 impeded the proliferation and migration and induced apoptosis of ESCC cells by regulating miR-498/VEGFA axis, which might serve as a novel biomarker for the diagnosis and treatment of ESCC.
Keywords: lncRNA AFAP1-AS1, miR-498, VEGFA, ESCC
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