Differential expression of circRNAs in anterior lens capsules of high myopic patients with cataract

doi:10.12122/j.issn.1673-4254.2025.09.19

Abstract

Abstract:

Objective To analyze the differential expression and biological functions of circRNAs in the anterior lens capsules of high myopic patients with cataract and their pathogenic roles in the development of this condition. Methods Anterior lens capsule specimens were collected intraoperatively from 36 patients with age-related cataract (ARC) and 36 high myopic patients with cataract. Among these, 18 specimens from each group were selected for whole transcriptome sequencing and biological analysis, and the remaining 36 specimens were used for validation of circPDGFRA, circFOXJ3, hsa_circ_0004767, hsa_circ_0007528, ciCRIM1, circMAN1A2, circSLC5A3, and circPTK2 expressions using RT-qPCR. hsa_circ_0007528 was selected for cell experiments to examine its effects on proliferation, migration, and apoptosis of lens epithelial cells (LECs). Results A total of 16 192 circRNAs were detected in the specimens from both groups, among which 62 circRNAs were differentially expressed (29 upregulated and 33 downregulated). GO and KEGG analyses revealed that the differentially expressed circRNAs were primarily localized in the cytoplasm, nucleoplasm, and endoplasmic reticulum, and were involved in signaling pathways associated with Gap junction and the PI3K-Akt, NF-κB, Jak-STAT, HIF-1, and MAPK signaling pathways. The ceRNA network predicted multiple target genes. RT-qPCR validation results were consistent with the sequencing data. In the LECs, upregulation of hsa_circ_0007528 significantly inhibited cell proliferation and migration and obviously promoted cell apoptosis. Conclusion The expression profile of circRNAs in the anterior lens capsule of high myopic patients with cataract differs from that of ARC patients. Upregulation of hsa_circ_0007528 inhibits LEC proliferation and migration and promotes cell apoptosis.

Key words: high myopia with cataract, circRNA spectrum, whole transcriptome sequencing, hsa_circ_0007528, lens epithelial cells

Yuanyuan HAN, Feng SUN, Yan LIU, Mengyue XU, Che XU, Na LI, Juan LI, Jianfeng WANG. Differential expression of circRNAs in anterior lens capsules of high myopic patients with cataract[J]. Journal of Southern Medical University, 2025, 45(9): 1997-2005.

Figures/Tables 6

Tab.1 Sequences of the primers for RT-qPCR of the circRNAs

circRNA	Forward (5'-3')	Reverse (5'-3')
ciRNA374	TGGGAATAGAACTGCTGA	ATGGGCTGCTTTGTAGAG
circRNA4906	ATTCCTTGGGCAATGGTG	GTGGCAGTGGCTTGTTCT
circRNA4418	GCATTAGCCTTCAAAAGT	CCACAAGACCATCAGCAT
circRNA4927	TCAGGGTGAAGAGCGATTA	CCATTGCACCAGGAGAAC
circRNA10422	TCTGCCTGACATTGACCC	TGAGCCTAACATAGATTTCTTC
circRNA12432	CAGCCTCATTACATTTGC	ATTTGGGTCAAGGAGTGC
hsa_circ_0007528	GCTTTGATCTTGCCTCTG	ATGCTGAACAATCGCTGA
hsa_circ_0004767	AAAATAGCCCTGAGTGAA	AACCTGATGCCTGAAGAC

Fig.1 Total circRNAs detected in the anterior capsular membranes of high myopia with cataract (HMC) and age-related cataract (ARC) lenses. A: Venn diagram of circRNAs in HMC group and ARC group. B: Volcano plot of differentially expressed circRNAs in the two groups (red represents up-regulated significantly differentially expressed genes, blue represents down-regulated ones, and gray represents those without significant differential expression. C: Bar chart of differentially expressed circRNAs. D: Proportional distribution chart of circRNA gene composition types. E: Length distribution chart of circRNAs. F: Clustering analysis chart of circRNAs: different colors represent the levels of expression (red represents high expression genes, and blue represents low expression genes).

Fig.2 GO and KEGG enrichment analysis of the differentially expressed circRNAs. A, C: Biological process, cellular component, and molecular function analysis of differentially up-regulated and down-regulated circRNAs. P<0.05; B, D: KEGG pathways of differentially up-regulated and down-regulated circRNAs, where Rich factor represents the number of differentially expressed genes in the KEGG pathway divided by the total number of genes in the KEGG pathway (Rich factor=S gene number/B gene number). The larger the Rich factor, the higher the degree of KEGG enrichment. P<0.05 indicates significant enrichment.

Fig.3 The ceRNA network diagram of circRNAs interaction. Circles represent circRNAs, arrows represent miRNAs, rectangles represent mRNAs, blue represents ceRNA interactions of up-regulated circRNAs, and red represents ceRNA interactions of down-regulated circRNAs.

Fig.4 Results of the RT-qPCR validation of the differentially expressed circRNAs. A: Expression of the up-regulated circRNAs in the anterior capsular tissue of the lens of HMC patients. B: Expression of the down-regulated circRNA in the anterior capsular tissue of the lens of HMC patients. **P<0.01, ***P< 0.001, ****P<0.0001.

Fig.5 Overexpression of hsa_circ_0007528 inhibits proliferation and migration of LECs and promotes apoptosis. A: Transfection efficiency of hsa_circ_0007528 verified by RT-qPCR. B: CCK-8 assay for assessing cell proliferation of LECs with hsa_circ_0007528 overexpression. C: Transwell cell migration assay for assessing changes in migration of LECs with hsa_circ_0007528 overexpression (Scale bar=50 μm). D: Flow cytometry for analyzing changes in apoptosis of LECs with hsa_circ_0007528 overexpression. n=3, **P<0.01, ***P<0.001, ****P<0.0001.

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