LncRNA AFAP1-AS1/miR-27b-3p/VEGF-C axis modulates stemness characteristics in cervical cancer cells.
10.1097/CM9.0000000000001665
- Author:
Meng XIA
1
;
Li-Jun DUAN
2
;
Bi-Nan LU
1
;
Yu-Zhou PANG
3
;
Zong-Ran PANG
1
Author Information
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.
- Publication Type:Journal Article
- MeSH:
Cell Line, Tumor;
Cell Proliferation/genetics*;
Female;
Gene Expression Regulation, Neoplastic;
Humans;
MicroRNAs/genetics*;
RNA, Long Noncoding/genetics*;
Uterine Cervical Neoplasms/genetics*;
Vascular Endothelial Growth Factor A/genetics*;
Vascular Endothelial Growth Factor C
- From:
Chinese Medical Journal
2021;134(17):2091-2101
- CountryChina
- Language:English
-
Abstract:
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(+) CC cells had remarkable stemness and a high level of AFAP1-AS1. However, AFAP1-AS1 knockdown with siAFAP1-AS1 suppressed the cell cycle transition of G(1)/S phase and inhibited self-renewal of CD44v6(+) CC cells, the levels of the stemness markers octamer-binding transcription factor 4 (OCT4), osteopontin (OPN), and cluster of differentiation 133 (CD133), and the epithelial-mesenchymal 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.
