CircMYCBP2促进膀胱癌淋巴管癌栓形成的机制

doi:10.3969/j.issn.1006-5725.2025.08.008

Abstract

Abstract:

Objective To investigate the role of circular RNA， circMYCBP2， which is highly expressed in lymph node （LN） metastatic bladder cancer （BCa） tissues， in enhancing BCa cell adhesion and mediating lymphovascular invasion （LVI）， and to evaluate its clinical relevance and potential therapeutic value. Methods High-throughput sequencing identified circRNAs with high expression in LN metastatic BCa tissues. Further validation of their expression in bladder cancer tissues was conducted through clinical samples. The coding ability of circMYCBP2 was predicted by circBank bioinformatics website， and the encoded short peptide was detected by immunoprecipitation（co-IP）combined with silver staining and Western blot （WB）. Then， by constructing BCa cells with overexpression and knockdown of circMYCBP2 in vitro，the invasive ability of BCa cells was verified by wound healing and transendothelial cell migration assays. The underlying mechanism of circMYCBP2 in the formation of LVI was explored through RNA pulldown， qRT-PCR and WB. Results High-throughput sequencing and clinical sample validation confirmed that circMYCBP2 is highly expressed in LN metastatic BCa.In vitro experiments demonstrated that circMYCBP2 overexpression significantly enhanced BCa cell migration across lymphatic endothelial cells. Mechanistically，circMYCBP2 encodes the short peptide MYCBP2-227aa via an IRES-dependent mechanism by interacting with EIF3H.Overexpression of MYCBP2-227aa increased the mRNA stability of VCAM-1， thereby enhancing the invasive capacity of UM-UC-3 cells. Conclusions CircMYCBP2 encodes the short peptide MYCBP2-227aa through an EIF3H-mediated， IRES-dependent translation mechanism.MYCBP2-227aa regulates VCAM-1 expression and promotes the invasive behavior of BCa cells. Our findings elucidate the critical biological role and molecular mechanism of circMYCBP2 in the formation of LVI and LN metastasis of BCa， providing a potential biomarker and therapeutic target for early lymphatic metastasis in BCa.

Key words: bladder cancer, lymphovascular invasion, circRNA translation, IRES

CLC Number:

R694.7

Daiyin LIU,Qiuyan LI,Changhao. CHEN. CircMYCBP2 enhances the formation of lymphovascular invasion of bladder cancer[J]. The Journal of Practical Medicine, 2025, 41(8): 1139-1148.

Figures/Tables 7

Tab.1

Tab.2

Fig.1

Fig.2

Fig.3

Fig.4

Fig.5

References 34

1	ZHENG H， AN M， LUO Y， et al. PDGFRα（+）ITGA11（+） fibroblasts foster early-stage cancer lymphovascular invasion and lymphatic metastasis via ITGA11-SELE interplay ［J］. Cancer Cell， 2024， 42（4）： 682-700.e12. doi:10.1016/j.ccell.2024.02.002 doi: 10.1016/j.ccell.2024.02.002
2	LIN Y， ZHENG H， JIA L， et al. Integrin α6-containing extracellular vesicles promote lymphatic remodelling for pre-metastatic niche formation in lymph nodes via interplay with CD151 ［J］. J Extracell Vesicles， 2024， 13（10）： e12518. doi:10.1002/jev2.12518 doi: 10.1002/jev2.12518
3	LIU S， CHEN X， LIN T， et al. Lymphatic metastasis of bladder cancer： Molecular mechanisms， diagnosis and targeted therapy ［J］. Cancer Lett， 2021， 505： 13-23. doi:10.1016/j.canlet.2021.02.010 doi: 10.1016/j.canlet.2021.02.010
4	CHEN C， ZHENG H， LUO Y， et al. SUMOylation promotes extracellular vesicle-mediated transmission of lncRNA ELNAT1 and lymph node metastasis in bladder cancer ［J］. J Clin Invest， 2021， 131（8）：e146431. doi:10.1172/jci146431 doi: 10.1172/jci146431
5	GARCíA-SILVA S， BENITO-MARTÍN A， NOGUÉS L， et al. Melanoma-derived small extracellular vesicles induce lymphangiogenesis and metastasis through an NGFR-dependent mechanism ［J］. Nat Cancer， 2021， 2（12）： 1387-1405. doi:10.1038/s43018-021-00272-y doi: 10.1038/s43018-021-00272-y
6	MORI K， SCHUETTFORT V M， KATAYAMA S， et al. Prognostic Role of Preoperative Vascular Cell Adhesion Molecule-1 Plasma Levels in Urothelial Carcinoma of the Bladder Treated With Radical Cystectomy ［J］. Ann Surg Oncol， 2022， 29（8）： 5307-5316. doi:10.1245/s10434-022-11575-4 doi: 10.1245/s10434-022-11575-4
7	ZHANG Q， LIU S， WANG H， et al. ETV4 Mediated Tumor-Associated Neutrophil Infiltration Facilitates Lymphangiogenesis and Lymphatic Metastasis of Bladder Cancer ［J］. Adv Sci （Weinh）， 2023， 10（11）： e2205613. doi:10.1002/advs.202370065 doi: 10.1002/advs.202370065
8	KOINA M E， BAXTER L， ADAMSON S J， et al. Evidence for lymphatics in the developing and adult human choroid ［J］. Invest Ophthalmol Vis Sci， 2015， 56（2）： 1310-1327. doi:10.1167/iovs.15-18011 doi: 10.1167/iovs.15-18011
9	FAN X， YANG Y， CHEN C， et al. Pervasive translation of circular RNAs driven by short IRES-like elements ［J］. Nat Commun， 2022， 13（1）： 3751. doi:10.1038/s41467-022-31327-y doi: 10.1038/s41467-022-31327-y
10	TAKALLOU S， PUCHACZ N， ALLARD D， et al. IRES-mediated translation in bacteria ［J］. Biochem Biophys Res Commun， 2023， 641： 110-115. doi:10.1016/j.bbrc.2022.12.022 doi: 10.1016/j.bbrc.2022.12.022
11	CHEN C K， CHENG R， DEMETER J， et al. Structured elements drive extensive circular RNA translation ［J］. Mol Cell， 2021， 81（20）： 4300-4318.e13. doi:10.1016/j.molcel.2021.07.042 doi: 10.1016/j.molcel.2021.07.042
12	LI Y， WANG Z， SU P， et al. circ-EIF6 encodes EIF6-224aa to promote TNBC progression via stabilizing MYH9 and activating the Wnt/beta-catenin pathway ［J］. Mol Ther， 2022， 30（1）： 415-430. doi:10.1016/j.ymthe.2021.08.026 doi: 10.1016/j.ymthe.2021.08.026
13	BEHRINGER A， STOIMENOVSKI D， PORSCH M， et al. Relationship of micro-RNA， mRNA and eIF Expression in Tamoxifen-Adapted MCF-7 Breast Cancer Cells： Impact of miR-1972 on Gene Expression， Proliferation and Migration ［J］. Biomolecules， 2022， 12（7）：916. doi:10.3390/biom12070916 doi: 10.3390/biom12070916
14	XU Y， GAO Z， SUN X， et al. The role of circular RNA during the urological cancer metastasis： Exploring regulatory mechanisms and potential therapeutic targets ［J］. Cancer Metastasis Rev， 2024， 43（3）： 1055-1074. doi:10.1007/s10555-024-10182-x doi: 10.1007/s10555-024-10182-x
15	AN M， ZHENG H， HUANG J， et al. Aberrant Nuclear Export of circNCOR1 Underlies SMAD7-Mediated Lymph Node Metastasis of Bladder Cancer ［J］. Cancer Res， 2022， 82（12）： 2239-2253. doi:10.1158/0008-5472.can-21-4349 doi: 10.1158/0008-5472.can-21-4349
16	PAN X， CHEN K， GAO W， et al. Circular RNA circBNC2 inhibits tumorigenesis by modulating ferroptosis and acts as a nanotherapeutic target in prostate cancer ［J］. Mol Cancer， 2025， 24（1）： 29. doi:10.1186/s12943-025-02234-9 doi: 10.1186/s12943-025-02234-9
17	YANG X， YE T， LIU H， et al. Expression profiles， biological functions and clinical significance of circRNAs in bladder cancer ［J］. Mol Cancer， 2021， 20（1）： 4. doi:10.1186/s12943-020-01300-8 doi: 10.1186/s12943-020-01300-8
18	MARQUES R， LACERDA R， ROMÃO L， et al. Internal Ribosome Entry Site （IRES）-Mediated Translation and Its Potential for Novel mRNA-Based Therapy Development ［J］. Biomedicines， 2022， 10（8）：1865. doi:10.3390/biomedicines10081865 doi: 10.3390/biomedicines10081865
19	HWANG H J， KIM Y K. Molecular mechanisms of circular RNA translation ［J］. Exp Mol Med， 2024， 56（6）： 1272-1280. doi:10.1038/s12276-024-01220-3 doi: 10.1038/s12276-024-01220-3
20	CHEN L， WANG C， SUN H， et al. The bioinformatics toolbox for circRNA discovery and analysis ［J］. Brief Bioinform， 2021， 22（2）： 1706-1728. doi:10.1093/bib/bbaa001 doi: 10.1093/bib/bbaa001
21	ZHAO X， ZHONG Y， WANG X， et al. Advances in Circular RNA and Its Applications ［J］. Int J Med Sci， 2022， 19（6）： 975-985. doi:10.7150/ijms.71840 doi: 10.7150/ijms.71840
22	LUíS C， SOARES R， FERNANDES R， et al. Cell-adhesion Molecules as Key Mechanisms of Tumor Invasion： The Case of Breast Cancer ［J］. Curr Mol Med， 2023， 23（2）： 147-160. doi:10.2174/1566524021666210806155231 doi: 10.2174/1566524021666210806155231
23	SMART J A， OLEKSAK J E， HARTSOUGH E J， et al. Cell Adhesion Molecules in Plasticity and Metastasis ［J］. Mol Cancer Res， 2021， 19（1）： 25-37. doi:10.1158/1541-7786.mcr-20-0595 doi: 10.1158/1541-7786.mcr-20-0595
24	陈立智，胡军，傅厚丰. L1细胞黏附分子在结肠癌中表达的临床意义和生物学作用［J］. 实用医学杂志， 2020， 36（19）： 2690-2695.
25	BUI T M， WIESOLEK H L， SUMAGIN R， et al. ICAM-1： A master regulator of cellular responses in inflammation， injury resolution， and tumorigenesis ［J］. J Leukoc Biol， 2020， 108（3）： 787-799. doi:10.1002/jlb.2mr0220-549r doi: 10.1002/jlb.2mr0220-549r
26	BASSIOUNI W， ALI M A M， SCHULZ R， et al. Multifunctional intracellular matrix metalloproteinases： Implications in disease ［J］. FEBS J， 2021， 288（24）： 7162-7182. doi:10.1111/febs.15701 doi: 10.1111/febs.15701
27	王子文，赵文静，晁亚丽. D-二聚体，乳酸联合可溶性血小板内皮黏附分子-1对脓毒症相关弥散性血管内凝血患者预后不良的预测研究［J］. 实用医学杂志， 2023， 39（18）： 2379-2383.
28	POWER G， FERREIRA-SANTOS L， MARTINEZ-LEMUS L A， et al. Integrating molecular and cellular components of endothelial shear stress mechanotransduction ［J］. Am J Physiol Heart Circ Physiol， 2024， 327（4）： H989-H1003. doi:10.1152/ajpheart.00431.2024 doi: 10.1152/ajpheart.00431.2024
29	赵莉莉，杨筱青，张瑞丽. 妊娠期糖尿病患者肿瘤坏死因子-α和血管细胞黏附分子-1的表达与母婴结局的关系［J］. 实用医学杂志， 2020， 36（13）： 1793-1796.
30	VANHEYST K A， CHOI S H， KINGSLEY D T， et al. Ectopic Tumor VCAM-1 Expression in Cancer Metastasis and Therapy Resistance ［J］. Cells， 2022， 11（23）：3922. doi:10.3390/cells11233922 doi: 10.3390/cells11233922
31	吴田方，戴东，王常元. circ-0008583结合EIF4A3蛋白促进肝癌细胞增殖并抑制其凋亡［J］. 实用医学杂志， 2022， 38（9）： 1088-1093 . doi:10.3969/j.issn.1006-5725.2022.09.009 doi: 10.3969/j.issn.1006-5725.2022.09.009
32	吴文松，刘超，孙程圆. circRNA在胶质瘤中的研究进展［J］. 实用医学杂志， 2021， 37（18）： 2317-2321 .
33	XIE B， LIN J， CHEN X， et al. CircXRN2 suppresses tumor progression driven by histone lactylation through activating the Hippo pathway in human bladder cancer ［J］. Mol Cancer， 2023， 22（1）： 151. doi:10.1186/s12943-023-01856-1 doi: 10.1186/s12943-023-01856-1
34	SHI X， PANG S， ZHOU J， et al. Bladder-cancer-derived exosomal circRNA_0013936 promotes suppressive immunity by up-regulating fatty acid transporter protein 2 and down-regulating receptor-interacting protein kinase 3 in PMN-MDSCs ［J］. Mol Cancer， 2024， 23（1）： 52. doi:10.1186/s12943-024-01968-2 doi: 10.1186/s12943-024-01968-2

引物名称	序列（5′-3′）
circMYCBP2上游引物	GAGATGTCAACTCCAGGGGA
circMYCBP2下游引物	CAGCACAACCAGATTCTCCG
GAPDH上游引物	CACATCGCTCAGACACCATG
GAPDH下游引物	TGACGGTGCCATGGAATTTG
VCAM-1上游引物	AAGGTTCCTAGCGTGTACCC
VCAM-1下游引物	CTGCCTTTGTTTGGGTTCGA
U1上游引物	CAGGGGAGATAACGTGACCA
U1下游引物	GGGAAAAGCACGGACACAG
18S rRNA上游引物	AGCAGACATTGACCTCACCA
18S rRNA下游引物	CCTCTATGGGCCCGAATCTT
si-has_circ_0004217正义链	UUUGGAAGUUUUUCUGAUCUGTT
si-has_circ_0004217反义链	CAGAUCAGAAAAACUUCCAAATT
si-EIF3H-1正义链	GGUAUUAAAGAUAAUCAAATT
si-EIF3H-1反义链	UUUGAUUAUCUUUAAUACCTT
si-EIF3H-2正义链	CAAACAUUAUCAAGAAGAATT
si-EIF3H-2反义链	UUCUUCUUGAUAAUGUUUGTT
si-EIF3H-3正义链	GAUGGAAGUUUGUAAAGAATT
si-EIF3H-3反义链	UUCUUUACAAACUUCCAUCTT

名称	公司	货号
Anti-EIF3H	Immunoway	YN3058
Anti-MYCBP2-227aa	杭州华安生物	RB4657

[1]	Yawei ZHANG,Hongjin SHI,Shi FU,Jiansong WANG,Haifeng WANG. Research progress in biological role of TIGIT and its application in bladder cancer [J]. The Journal of Practical Medicine, 2024, 40(12): 1762-1766.
[2]	Feng XIAO,Xi′nan JIANG,Lu SHU,Jiaqi SHI,Fangmin CHEN,Yi. MU. Expression of ECM1 gene in bladder cancer and its relationships with clinical pathological features and prognosis [J]. The Journal of Practical Medicine, 2023, 39(20): 2629-2632.
[3]	Kai DENG,Meng YANG,Ling ZHANG,Jun′an QIAN,Yunqiang SHI,Chunhui. WANG. Bidirectional regulation of tumor associated neutrophils and their role in urothelial bladder cancer： a literature review [J]. The Journal of Practical Medicine, 2023, 39(16): 2142-2147.
[4]	YANG Wenbo, XU Zhendan, GENG Shuai. Effect of LINC00689 on proliferation and apoptosis of bladder cancer T24 cells through targeting miR⁃876⁃ 5p [J]. The Journal of Practical Medicine, 2021, 37(2): 195-200.

CircMYCBP2 enhances the formation of lymphovascular invasion of bladder cancer

RichHTML

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

Figures/Tables 7

References 34

Related Articles 4

Recommended Articles

Metrics

Comments