高度近视白内障患者晶状体前囊膜中circRNAs的差异性表达

doi:10.12122/j.issn.1673-4254.2025.09.19

摘要/Abstract

摘要：

目的分析高度近视白内障（HMC）患者晶状体前囊膜组织中circRNAs的差异性表达及其生物学功能，探讨circRNA在高度近视白内障中的作用。方法通过白内障手术术中获取36例年龄相关性白内障（ARC）和36例HMC患者晶状体前囊膜标本，两组中各选取18例进行全转录组测序和生物学分析。余下36例采用RT-qPCR对circPDGFRA、circFOXJ3、hsa_circ_0004767、hsa_circ_0007528、ciCRIM1、circMAN1A2、circSLC5A3、circPTK2进行临床验证。随后选择hsa_circ_0007528进行细胞实验，检测hsa_circ_0007528对晶状体上皮细胞（LECs）增殖、迁移和凋亡的影响。结果两组患者标本中共检测出16192个circRNAs，其中62个circRNAs差异性表达（29个上调、33个下调）。GO和KEGG分析检测，差异性表达的circRNAs多集中在细胞质、核质和内质网中，参与Gap junction、PI3K-Akt signaling pathway、NF-kappa B signaling pathway、Jak-STAT signaling pathway、HIF-1 signaling pathway、MAPK signaling pathway等信号通路（P<0.05）。ceRNA网络预测多个靶基因。RT-qPCR验证与测序结果一致（P<0.01）。CCK-8、Transwell结果显示，hsa_circ_0007528上调抑制了细胞增殖（P<0.001）和迁移（P<0.01）。流式细胞术显示hsa_circ_0007528上调促进了细胞的凋亡（P<0.0001）。结论 HMC患者晶状体前囊膜中circRNAs的表达谱与ARC存在差异。hsa_circ_0007528上调抑制LECs的增殖、迁移、促进细胞凋亡。

关键词: 高度近视白内障, circRNA谱, 全转录组测序, hsa_circ_0007528, 晶状体上皮细胞

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

韩圆圆, 孙凤, 刘艳, 徐梦月, 许澈, 李娜, 李娟, 王剑锋. 高度近视白内障患者晶状体前囊膜中circRNAs的差异性表达[J]. 南方医科大学学报, 2025, 45(9): 1997-2005.

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.

图/表 6

表1 用于RT-qPCR验证的circRNAs的引物序列

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

图1 HMC和ARC晶状体前囊膜中检测的总circRNAs

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).

图2 差异性表达circRNAs的GO和KEGG富集分析

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.

图3 差异性表达circRNAs的ceRNA网络

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.

图4 差异性表达的circRNAs的RT-qPCR验证结果

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.

图5 过表达hsa_circ_0007528抑制LECs的增殖和迁移,促进细胞凋亡

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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