LncRNA AFAP1-AS1/miR-27b-3p/VEGF-C axis modulates stemness characteristics in cervical cancer cells  

作　　者：Meng Xia Li-Jun Duan Bi-Nan Lu Yu-Zhou Pang Zong-Ran Pang 

机构地区：[1]School of Pharmacy,Minzu University of China,Beijing 100081,China [2]Department of Orthopedics,Bayannaoer City Hospital,Bayannaoer,Inner Mongolia 015000,China [3]Guangxi Zhuang Yao Medicine Center of Engineering and Technology,Guangxi University of Chinese Medicine,Nanning,Guangxi 530200,China [4]Key Laboratory of Ethnomedicine of Ministry of Education,Minzu University of China,Beijing 100081,China

出　　处：《Chinese Medical Journal》2021年第17期2091-2101,共11页中华医学杂志（英文版）

摘　　要：Background:Long non-coding RNA(lncRNA)actin filament-associated protein 1 antisense RNA 1(AFAP1-AS1)functions as a competing endogenous RNA to regulate target genes expression by sponging microRNAs(miRs)to play cancer-promoting roles in cancer stem cells.However,the regulatory mechanism of AFAP1-AS1 in cervical cancer(CC)stem cells is unknown.The present study aimed to provide a new therapeutic target for the clinical treatment of CC.Methods:Hyaluronic acid receptor cluster of differentiation 44 variant exon 6(CD44v6)(+)CC cells were isolated by flow cytometry(FCM).Small interfering RNAs of AFAP1-AS1(siAFAP1-AS1)were transfected into the(CD44v6)(+)cells.The levels of AFAP1-AS1 were measured by quantitative real-time PCR(qRT-PCR).Sphere formation assay,cell cycle analysis,and Western blotting were used to detect the effect of siAFAP1-AS1.RNA pull-down and luciferase reporter assay were used to verify the relationship between miR-27b-3p and AFAP1-AS1 or vascular endothelial growth factor(VEGF)-C.Results:CD44v6(+)CCcells had remarkable stemness and a high level ofAFAP1-AS1.However,AFAP1-AS1knockdownwithsiAFAP1-AS1suppressed the cell cycle transitionofG(1)/S phase and inhibited self-renewal ofCD44v6(+)CCcells,the levels of the stemnessmarkers octamer-binding transcription factor 4(OCT4),osteopontin(OPN),and cluster of differentiation 133(CD133),and the epithelialmesenchymal transition(EMT)-related proteins Twist1,matrix metalloprotease(MMP)-9,and VEGF-C.In the mechanism study,miR-27b-3p/VEGF-C signaling was demonstrated to be a key downstream of AFAP1-AS1 in the CD44v6(+)CC cells.Conclusions:LncRNA AFAP1-AS1 knockdown inhibits the CC cell stemness by upregulating miR-27b-3p to suppress VEGF-C.

关 键 词：Hyaluronic acid receptor cluster of differentiation 44 variant exon 6 Cell stemness Cervical cancer Long non-coding RNA actin filament-associated protein 1 antisense RNA 1 MicroRNA-27b-3p 

分 类 号：R737.33[医药卫生—肿瘤]