Circular RNA circ-FAM158A promotes retinoblastoma progression by regulating miR-138-5p/SLC7A5 axis

Tongmei Zheng; Weifang Chen; Xiuchun Wang; Weiguo Cai; Fujin Wu; Chaobin Lin

doi:10.1016/j.exer.2021.108650

Circular RNA circ-FAM158A promotes retinoblastoma progression by regulating miR-138-5p/SLC7A5 axis

Exp Eye Res. 2021 Oct:211:108650. doi: 10.1016/j.exer.2021.108650. Epub 2021 Jun 5.

Authors

Tongmei Zheng¹, Weifang Chen¹, Xiuchun Wang¹, Weiguo Cai¹, Fujin Wu¹, Chaobin Lin²

Affiliations

¹ Department of Ophthalmology, The First Hospital of Quanzhou Affiliated to Fujian Medical University, No.250 East Street, Quanzhou 362000, Fujian Province, China.
² Department of Ophthalmology, The First Hospital of Quanzhou Affiliated to Fujian Medical University, No.250 East Street, Quanzhou 362000, Fujian Province, China. Electronic address: ztm120627@163.com.

PMID: 34102206
DOI: 10.1016/j.exer.2021.108650

Abstract

Background: Mounting evidence has shown that circular RNAs (circRNAs) have vital roles in human cancers, including retinoblastoma (RB). The purpose of this study was to investigate the exact roles and underlying mechanism of circRNA ER membrane protein complex subunit 9 (circ-FAM158A) in RB.

Methods: Quantitative real-time polymerase chain reaction (qRT-PCR) was carried out to determine the expression levels of circ-FAM158A, miR-138-5p and solute carrier family 7 member 5 (SLC7A5). Cell proliferation was evaluated by Cell counting Kit-8 (CCK-8) assay and colony formation assay. Flow cytometry analysis was applied to determine cell cycle distribution and apoptosis rate. Transwell assay was conducted to assess cell migration and invasion. The interaction between miR-138-5p and circ-FAM158A or SLC7A5 was predicted by starBase v2.0 and confirmed by dual-luciferase reporter assay. Western blot assay was performed to examine the protein expression of SLC7A5. The mice xenograft model was established, immunohistochemistry (IHC) and terminal deoxynucleotidyl transferase-mediated dUTP nick end-labelling (TUNEL) assays were conducted to confirm the role of circ-FAM158A in vivo.

Results: Circ-FAM158A and SLC7A5 were overexpressed and miR-138-5p was downregulated in RB tissues and cells. Circ-FAM158A knockdown inhibited RB cell proliferation, metastasis, and promoted apoptosis in vitro and in vivo. MiR-138-5p was a direct target of circ-FAM158A, and miR-138-5p inhibition reversed the inhibitory effect of circ-FAM158A silence on the progression of RB cells. Additionally, SLC7A5 was identified as a target of miR-138-5p, and SLC7A5 overexpression abated the anti-tumor roles of miR-138-5p in RB cells. Besides, circ-FAM158A positively regulated SLC7A5 expression by sponging miR-138-5p.

Conclusion: Circ-FAM158A knockdown inhibited the progression of RB by regulating miR-138-5p/SLC7A5 axis, which provided new insights into the pathogenesis of RB.

Keywords: Circ-FAM158A; Retinoblastoma; SLC7A5; miR-138–5p.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Apoptosis
Blotting, Western
Cell Movement
Cell Proliferation
Disease Progression
Female
Flow Cytometry
Gene Expression Regulation, Neoplastic / physiology*
Gene Silencing
Heterografts
Humans
Immunohistochemistry
In Situ Nick-End Labeling
Large Neutral Amino Acid-Transporter 1 / genetics*
Membrane Proteins / genetics*
Mice
Mice, Inbred BALB C
Mice, Nude
MicroRNAs / genetics*
RNA, Circular / genetics*
Real-Time Polymerase Chain Reaction
Retinal Neoplasms / genetics*
Retinal Neoplasms / pathology
Retinoblastoma / genetics*
Retinoblastoma / pathology
Transfection
Tumor Cells, Cultured
Up-Regulation

Substances

Large Neutral Amino Acid-Transporter 1
MIRN138 microRNA, human
Membrane Proteins
MicroRNAs
RNA, Circular
TMEM9 protein, human