LncRNA AFAP1-AS1 Modulates the Proliferation and Invasion of Gastric Cancer Cells by Regulating AFAP1 via miR-205-5p

Abstract

Yuan Dang,1 Xiaojuan Ouyang,2 Wenjun Ren,3 Lie Wang,3 Qiaojia Huang1 1Laboratory of Basic Medicine, 900 Hospital of the Joint Logistics Team (Dongfang Hospital)(Former Fuzhou General Hospital), Xiamen University Medical College, Fuzhou, 350025, Fujian, People’s Republic of China; 2Department of Department of Pathology, 900 Hospital of the Joint Logistics Team (Dongfang Hospital) (Former Fuzhou General Hospital), Fuzhou, 350025, Fujian, People’s Republic of China; 3Department of General Surgery, 900 Hospital of the Joint Logistics Team (Dongfang Hospital) (Former Fuzhou General Hospital), Fuzhou, 350025, Fujian, People’s Republic of ChinaCorrespondence: Lie WangDepartment of General Surgery, 900 Hospital of the Joint Logistics Team (Dongfang Hospital) (Former Fuzhou General Hospital), 156 North Xi-er Huan Road, Fuzhou City, Fujian Province, 350025, People’s Republic of ChinaTel [phone]Fax [phone]Email [email protected] HuangLaboratory of Basic Medicine, 900 Hospital of the Joint Logistics Team (Dongfang Hospital) (Former Fuzhou General Hospital), 156 North Xi-er Huan Road, Fuzhou City, Fujian Province, 350025, People’s Republic of ChinaTel [phone]Fax [phone]Email [email protected]: The present study investigated the expression and function of the long noncoding RNA (lncRNA) actin filament associated protein 1 antisense RNA1 (AFAP1-AS1) related to gastric cancer (GC), based on previous results from a microarray analysis.Methods: Real-time quantitative polymerase chain reaction (qPCR) was used to verify the expression of AFAP1-AS1 in 97 fresh GC tissues and paired non-GC tissues, as well as in six different GC cell lines (BGC-823, SGC-7901, MGC-803, AGS, MKN-45, and MKN-28). The expression levels were subsequently correlated with the clinicopathological features of patients. siRNA against AFAP1-AS1 was transfected into GC cell lines, and cell proliferation, migration, and invasion were detected before and after silencing of AFAP1-AS1 expression. Luciferase reporter gene analysis was used to confirm the target gene of microRNA-205-5p (miR-205-5p) in 293T cells. The potential mechanism was subsequently investigated.Results: qPCR results showed that AFAP1-AS1 was significantly overexpressed in GC tumor tissues and also GC cell lines, comparing to their paired non-GC tissues. Furthermore, statistical analysis revealed that the overexpression of AFAP1-AS1 was significantly correlated with tumor size (p=0.018) and grade of differentiation (p=0.042). Subsequently, artificially decreasing the expression of AFAP1-AS1 with its specific siRNA dramatically inhibited the proliferation, migration and invasion of GC cell lines (SGC-7901 and BGC-823 cells). Mechanical analysis suggested that AFAP1-AS1 is involved in regulation of its maternal gene, AFAP1, at both mRNA level and protein level. Luciferase reporter gene assay indicated that lncRNA AFAP1-AS1, as a ceRNA, is able to sponge miR-205-5p. Moreover, miR-205-5p has been well demonstrated to participate in the regulation of AFAP1 expression and the phenotypes of GC cells, including proliferation, migration and invasion.Conclusion: AFAP1-AS1, as a novel biomarker of GC, promotes the proliferation migration and invasion of GC cells and function as ceRNA to target AFAP1 by sponging miR-205-5p.Keywords: gastric cancer, lncRNA, AFAP1-AS1, AFAP1, miR-205-5p

Keywords

• gastric cancer
• lncrna
• afap1-as1
• afap1
• mir-205-5p