hsa_circ_0001900 enhances the proliferation, migration, and invasion of Wilms tumor cells while suppressing apoptosis through acceleration of G1/S cell cycle transition

doi:10.3969/j.issn.1674-8115.2026.06.006

Abstract

Abstract:

Objective ·To explore the function and potential mechanism of circular RNA (circRNA) hsa_circ_0001900 in Wilms tumor. Methods ·Lentivirus was used to construct cell lines with stable overexpression of hsa_circ_0001900 (WIT-49, WT-CLS1, and SK-NEP-1), and specific small interfering RNA (siRNA) was used to transiently silence this molecule. Transcriptome sequencing was performed on hsa_circ_0001900-overexpressing and control cell lines, and its potential biological functions were preliminarily explored based on enrichment analysis of differentially expressed genes. cell counting kit-8 (CCK-8) assay, wound-healing assay, Transwell assay, and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay were used to evaluate the effects of hsa_circ_0001900 on the proliferation, migration, invasion, and apoptosis of Wilms tumor cells. Gene set enrichment analysis (GSEA) and gene set variation analysis (GSVA) were further adopted to screen Kyoto Encyclopedia of Genes and Genomes (KEGG) signaling pathways potentially regulated by hsa_circ_0001900. Finally, flow cytometry was used to detect cell cycle distribution and apoptosis to verify the related pathways experimentally. Results ·PCR assays confirmed that the expression of hsa_circ_0001900 was significantly upregulated in Wilms tumor cell lines (WIT-49, WT-CLS1, and SK-NEP-1) compared with normal human embryonic kidney HEK-293T cells. Cell lines with stable overexpression and transient silencing were successfully constructed using lentivirus and siRNA, respectively. Enrichment analysis of transcriptomic data showed that differentially expressed genes were mainly enriched in pathways associated with cell proliferation, cell cycle regulation, migration, and apoptosis. In vitro experiments further confirmed that overexpression of hsa_circ_0001900 promoted the proliferation, migration, and invasion of Wilms tumor cells, while silencing this molecule significantly inhibited the above malignant biological behaviors and induced cell apoptosis. Mechanistically, enrichment analysis centered on hsa_circ_0001900 revealed that GSEA and GSVA jointly enriched the cell cycle signaling pathway. Flow cytometry detection showed that stable overexpression of hsa_circ_0001900 markedly shortened the G1 phase and prolonged the S phase in WIT-49, WT-CLS1, and SK-NEP-1 cells, suggesting that this molecule may drive the malignant progression of Wilms tumor by facilitating G1/S phase transition. Conclusion ·hsa_circ_0001900 is specifically highly expressed in Wilms tumor cells. This molecule promotes the proliferation, migration, and invasion of Wilms tumor cells and inhibits cell apoptosis by accelerating the G1/S cell cycle transition.

Key words: Wilms tumor, circular RNA (circRNA), hsa_circ_0001900, cell cycle

CLC Number:

R726.9

Zhu Rundong, Gao Zhiqiang, Xiang Bin, Hong Peng, Hu Zaihong, Cui Kongkong, Long Teng, Zhang Zhijun, Ma Wei, Chen Pengfei, Shi Qinlin, Tian Xiaomao, Wei Guanghui. hsa_circ_0001900 enhances the proliferation, migration, and invasion of Wilms tumor cells while suppressing apoptosis through acceleration of G1/S cell cycle transition[J]. Journal of Shanghai Jiao Tong University (Medical Science), 2026, 46(6): 740-750.

Figures/Tables 8

Tab 1 siRNA sequences used for specific silencing of hsa_circ_0001900

siRNA	Target sequence(5'→3')
siRNA-1 (si-hsa_circ_0001900-1)	CTGGATCAACAAGATAACA
siRNA-2 (si-hsa_circ_0001900-2)	TCAACAAGATAACATCCCT
siRNA-3 (si-hsa_circ_0001900-3)	CAAGATAACATCCCTGAGG

Fig 1 Construction of Wilms tumor cell lines with stable overexpression of hsa_circ_0001900 using lentiviral Vectors

Fig 2 Transcriptome sequencing and bioinformatics analyses based on stably transfected cell lines

Fig 3 Overexpression of hsa_circ_0001900 promotes proliferation, migration, and invasion of Wilms tumor cells in vitro

Fig 4 Silencing hsa_circ_0001900 inhibits the proliferation, migration, and invasion of Wilms tumor cells

Fig 5 Silencing hsa_circ_0001900 induces apoptosis in WIT-49 Wilms tumor cells

Fig 6 GSEA, GSVA, and key mechanism prediction of hsa_circ_0001900 based on transcriptome sequencing

Fig 7 hsa_circ_0001900 regulates the cell cycle pathway in Wilms tumor cells by promoting G1/S phase transition

TrendMD

References 37

[1]	Sanger H L, Klotz G, Riesner D, et al. Viroids are single-stranded covalently closed circular RNA molecules existing as highly base-paired rod-like structures[J]. Proc Natl Acad Sci U S A, 1976, 73(11): 3852-3856.
[2]	Cocquerelle C, Mascrez B, Hétuin D, et al. Mis-splicing yields circular RNA molecules[J]. FASEB J, 1993, 7(1): 155-160.
[3]	Capel B, Swain A, Nicolis S, et al. Circular transcripts of the testis-determining gene Sry in adult mouse testis[J]. Cell, 1993, 73(5): 1019-1030.
[4]	Salzman J, Gawad C, Wang P L, et al. Circular RNAs are the predominant transcript isoform from hundreds of human genes in diverse cell types[J]. PLoS One, 2012, 7(2): e30733.
[5]	Ivanov A, Memczak S, Wyler E, et al. Analysis of intron sequences reveals hallmarks of circular RNA biogenesis in animals[J]. Cell Rep, 2015, 10(2): 170-177.
[6]	Salzman J, Chen R E, Olsen M N, et al. Cell-type specific features of circular RNA expression[J]. PLoS Genet, 2013, 9(9): e1003777.
[7]	Xia S Y, Feng J, Lei L J, et al. Comprehensive characterization of tissue-specific circular RNAs in the human and mouse genomes[J]. Brief Bioinform, 2017, 18(6): 984-992.
[8]	Memczak S, Jens M, Elefsinioti A, et al. Circular RNAs are a large class of animal RNAs with regulatory potency[J]. Nature, 2013, 495(7441): 333-338.
[9]	Fletcher K, Klosterman S J, Derevnina L, et al. Comparative genomics of downy mildews reveals potential adaptations to biotrophy[J]. BMC Genomics, 2018, 19(1): 851.
[10]	Gruner H, Cortés-López M, Cooper D A, et al. CircRNA accumulation in the aging mouse brain[J]. Sci Rep, 2016, 6: 38907.
[11]	Han B, Chao J, Yao H H. Circular RNA and its mechanisms in disease: from the bench to the clinic[J]. Pharmacol Ther, 2018, 187: 31-44.
[12]	Vo J N, Cieslik M, Zhang Y J, et al. The landscape of circular RNA in cancer[J]. Cell, 2019, 176(4): 869-881.e13.
[13]	Tian X M, Xiang B, Zhang Z X, et al. The regulatory network and role of the circRNA-miRNA-mRNA ceRNA network in the progression and the immune response of wilms tumor based on RNA-seq[J]. Front Genet, 2022, 13: 849941.
[14]	高志强, 林洁, 洪鹏, 等. 高通量环状RNA测序揭示hsa＿circ＿0001900在肾母细胞瘤中特异性高表达且与不良预后相关[J]. 南方医科大学学报, 2025, 45(11): 2466-2474.
	Gao Z Q, Lin J, Hong P, et al. High-throughput circular RNA sequencing reveals tumor-specific high expression of hsa_circ_0001900 in Wilms tumor in association with poor prognosis[J]. Journal of Southern Medical University, 2025, 45(11): 2466-2474.
[15]	Wang Y A, Yan Q J, Mo Y Z, et al. Splicing factor derived circular RNA circCAMSAP1 accelerates nasopharyngeal carcinoma tumorigenesis via a SERPINH1/c-Myc positive feedback loop[J]. Mol Cancer, 2022, 21(1): 62.
[16]	Luo Z J, Lu L B, Tang Q L, et al. CircCAMSAP1 promotes hepatocellular carcinoma progression through miR-1294/GRAMD1A pathway[J]. J Cell Mol Med, 2021, 25(8): 3793-3802.
[17]	Zhou C, Liu H S, Wang F W, et al. circCAMSAP1 promotes tumor growth in colorectal cancer via the miR-328-5p/E2F1 axis[J]. Mol Ther, 2020, 28(3): 914-928.
[18]	Buchfink B, Xie C, Huson D H. Fast and sensitive protein alignment using DIAMOND[J]. Nat Methods, 2015, 12(1): 59-60.
[19]	Robinson M D, McCarthy D J, Smyth G K. edgeR: a Bioconductor package for differential expression analysis of digital gene expression data[J]. Bioinformatics, 2010, 26(1): 139-140.
[20]	Subramanian A, Tamayo P, Mootha V K, et al. Gene set enrichment analysis: a knowledge-based approach for interpreting genome-wide expression profiles[J]. Proc Natl Acad Sci U S A, 2005, 102(43): 15545-15550.
[21]	Hänzelmann S, Castelo R, Guinney J. GSVA gene set variation analysis for microarray and RNA-seq data[J]. BMC Bioinform, 2013, 14: 7.
[22]	Ritchie M E, Phipson B, Wu D, et al. Limma powers differential expression analyses for RNA-sequencing and microarray studies[J]. Nucleic Acids Res, 2015, 43(7): e47.
[23]	Hwang H J, Kim Y K. Molecular mechanisms of circular RNA translation[J]. Exp Mol Med, 2024, 56(6): 1272-1280.
[24]	Zhang S B, Lin S Y, Liu M, et al. CircAnks1a in the spinal cord regulates hypersensitivity in a rodent model of neuropathic pain[J]. Nat Commun, 2019, 10: 4119.
[25]	Li H, Xu J D, Fang X H, et al. Circular RNA circRNA_000203 aggravates cardiac hypertrophy via suppressing miR-26b-5p and miR-140-3p binding to Gata4[J]. Cardiovasc Res, 2020, 116(7): 1323-1334.
[26]	Jiang Q, Liu C, Li C P, et al. Circular RNA-ZNF532 regulates diabetes-induced retinal pericyte degeneration and vascular dysfunction[J]. J Clin Invest, 2020, 130(7): 3833-3847.
[27]	Goodall G J, Wickramasinghe V O. RNA in cancer[J]. Nat Rev Cancer, 2021, 21(1): 22-36.
[28]	Liu Z H, Zhou Y, Liang G H, et al. Circular RNA hsa_circ_001783 regulates breast cancer progression via sponging miR-200c-3p[J]. Cell Death Dis, 2019, 10(2): 55.
[29]	Xie H J, Ren X L, Xin S N, et al. Emerging roles of circRNA_001569 targeting miR-145 in the proliferation and invasion of colorectal cancer[J]. Oncotarget, 2016, 7(18): 26680-26691.
[30]	Wang X L, Li J X, Bian X, et al. CircURI1 interacts with hnRNPM to inhibit metastasis by modulating alternative splicing in gastric cancer[J]. Proc Natl Acad Sci U S A, 2021, 118(33): e2012881118.
[31]	Rubin S M, Sage J, Skotheim J M. Integrating old and new paradigms of G1/S control[J]. Mol Cell, 2020, 80(2): 183-192.
[32]	Liu F, Qu R F, Yang L M, et al. Circular RNA controls tumor occurrence and development via cell cycle regulation[J]. Onco Targets Ther, 2022, 15: 993-1009.
[33]	Chao F, Wang S Y, Zhang C, et al. The emerging role of circular RNAs in prostate cancer: a systematic review[J]. Front Cell Dev Biol, 2021, 9: 681163.
[34]	Zhang N, Wang X J, Li Y, et al. Mechanisms and therapeutic implications of gene expression regulation by circRNA-protein interactions in cancer[J]. Commun Biol, 2025, 8(1): 77.
[35]	Kristensen L S, Andersen M S, Stagsted L V W, et al. The biogenesis, biology and characterization of circular RNAs[J]. Nat Rev Genet, 2019, 20(11): 675-691.
[36]	He A T, Liu J L, Li F Y, et al. Targeting circular RNAs as a therapeutic approach: current strategies and challenges[J]. Signal Transduct Target Ther, 2021, 6: 185.
[37]	Zhang Y, Xue W, Li X, et al. The biogenesis of nascent circular RNAs[J]. Cell Rep, 2016, 15(3): 611-624.