Expression profile of cellular microRNAs and their potential role in cervical cancer

Cover Page


Cite item

Full Text

Open Access Open Access
Restricted Access Access granted
Restricted Access Subscription or Fee Access

Abstract

Background. Cervical cancer (CC) is the fourth leading cause of cancer-related morbidity and mortality among women. Despite the established etiologic factor of cervical cancer, especially high-risk human papillomavirus (HPV, human papillomavirus) and available vaccine prophylaxis, the issue persists both in understanding the mechanism of HPV-associated carcinogenesis and in developing new approaches for diagnosis and treatment of cervical cancer. Epigenetic regulation of gene expression by means of microRNAs plays an important role in the pathogenesis of cervical cancer. Therefore, the search for new differentially expressed microRNAs in order to reveal new mechanisms of HPV-associated transformation of tumor cells is of high priority.

AIM: To explore microRNAs with differentiated expression in the cervical cancer tissue and to assess the functional potential of their detection in silico.

MATERIALS AND METHODS: The spectrum of microRNAs characterizing by modified expression in the tumor tissue of HPV16-positive squamous cell carcinomas of the cervix was determined using NGS sequencing (Next Generation Sequencing) of tumor tissue and of apparently unchanged adjacent epithelium obtained with the use of microdissection. Potential target genes of the investigated microRNAs were identified using MirTargetLink, Tarbase v9.0, and LinkedOmics services. Gene Set Enrichment Analysis was performed using Metascape. The relations between the level of microRNA expression and clinical SCC features (according to TCGA data, CESC sample) were searched using USCS Xena service.

RESULTS: NGS-sequencing of paired HPV16-positive specimens of tumor tissue and normal cervical tissue resulted in the identification of 42 differentially expressed microRNAs. Specifically, the levels of 22 microRNAs in the tumor tissue were higher than in the apparently normal adjacent epithelium, while 20 microRNAs demonstrated lower levels in the tumor specimens. Analysis of the potential targets of significant microRNAs revealed multiple functional gene categories, potentially involved in carcinogenesis as well as an association with clinical features. We showed that increased expression levels of microRNA-20b in the tumor tissue correlated with the risk of distant metastases, whereas the lower levels of microRNA-218-1 and microRNA -218-2 were associated with unfavorable prognosis of disease. With regard to microRNA-363, -615, and -769, their increase in cervical cancer was described for the first time, and the potential targets and signaling pathways associated with THEIR EXPRESSION LEVELS WERE IDENTIFIED.

CONCLUSION: The search for differentially expressed microRNAs in cervical cancer has revealed a spectrum of microRNAs with potentially important role in the process of malignant transformation and persistence of tumor phenotype. In addition to microRNAs demonstrating functional characteristics described in the world literature, we found microRNAs that play an unknown role in cervical cancer. In this relation, the potentially relevant targets were identified that could be helpful in understanding the mechanisms of carcinogenesis. The data obtained may form the basis for the development of new approaches to the diagnosis and therapy of SCC.

Full Text

Restricted Access

About the authors

Danila S. Elkin

Blokhin National Medical Research Center of Oncology

Author for correspondence.
Email: d.elkin@ronc.ru
ORCID iD: 0000-0002-4793-6063
SPIN-code: 9946-6863
Russian Federation, Moscow

Radik S. Faskhutdinov

Blokhin National Medical Research Center of Oncology

Email: r.faskhutdinov@ronc.ru
ORCID iD: 0000-0002-0050-7798
Russian Federation, Moscow

Olga V. Zvereva

Blokhin National Medical Research Center of Oncology

Email: zvereva.owl@mail.ru
ORCID iD: 0009-0007-7691-3235
Russian Federation, Moscow

Mariya D. Fedorova

Blokhin National Medical Research Center of Oncology

Email: m.d.fedorova@ronc.ru
ORCID iD: 0000-0002-8813-7516
SPIN-code: 4943-5931

Cand. Sci. (Biology)

Russian Federation, Moscow

Aleksej N. Katargin

Blokhin National Medical Research Center of Oncology

Email: a.katargin@ronc.ru
ORCID iD: 0000-0002-7405-0671

Cand. Sci. (Biology)

Russian Federation, Moscow

Larisa S. Pavlova

Blokhin National Medical Research Center of Oncology

Email: l.pavlova@ronc.ru
ORCID iD: 0000-0003-3993-4823
Russian Federation, Moscow

Kirill I. Zhordania

Blokhin National Medical Research Center of Oncology

Email: k.zhordania@ronc.ru
ORCID iD: 0000-0003-1380-3710
SPIN-code: 6271-8954

MD, Dr. Sci. (Medicine), Professor

Russian Federation, Moscow

Ekaterina A. Mustafina

Blokhin National Medical Research Center of Oncology

Email: e.mustafina@ronc.ru
ORCID iD: 0000-0002-1009-0383

MD, Cand. Sci. (Medicine)

Russian Federation, Moscow

Svetlana V. Vinokurova

Blokhin National Medical Research Center of Oncology

Email: s.vinokurova@ronc.ru
ORCID iD: 0000-0003-1615-3928
SPIN-code: 3453-4502

MD, Cand. Sci. (Medicine)

Russian Federation, Moscow

References

  1. Bray F, Laversanne M, Sung H, et al. Global cancer statistics 2022: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin. 2024;74(3):229–263. doi: 10.3322/caac.21834
  2. Egawa N, Egawa K, Griffin H, et al. Viruses. 2015;7(7):3863–3890. doi: 10.3390/v7072802
  3. Scarth J, Patterson M, Morgan E, Macdonald A. The human papillomavirus oncoproteins: a review of the host pathways targeted on the road to transformation. J Gen Virol. 2021;102(3): 001540. doi: 10.1099/jgv.0.001540
  4. Thierry F. Transcriptional regulation of the papillomavirus oncogenes by cellular and viral transcription factors in cervical carcinoma. Virology. 2009;384(2):375–379. doi: 10.1016/j.virol.2008.11.014
  5. Pett M, Coleman N. Integration of high-risk human papillomavirus: a key event in cervical carcinogenesis? J Pathol. 2007;212(4):356–367. doi: 10.1002/path.2192
  6. Vinokurova S, Wentzensen N, Kraus I, et al. Type-dependent integration frequency of human papillomavirus genomes in cervical lesions. Cancer Res. 2008;68(1):307–313. doi: 10.1158/0008-5472.CAN-07-2754
  7. Arias-Pulido H, Peyton C, Joste N, et al. Human papillomavirus type 16 integration in cervical carcinoma in situ and in invasive cervical cancer. J Clin Microbiol. 2006;44(5):1755–1762. doi: 10.1128/JCM.44.5.1755-1762.2006
  8. Klaes R, Friedrich T, Spitkovsky D, et al. Overexpression of p16(INK4A) as a specific marker for dysplastic and neoplastic epithelial cells of the cervix uteri. Int J Cancer. 2001;92(2):276–284. doi: 10.1002/ijc.1174
  9. Sano T, Oyama T, Kashiwabara K, et al. Expression status of p16 protein is associated with human papillomavirus oncogenic potential in cervical and genital lesions. Am J Pathol. 1998;153(6):1741–1748. doi: 10.1016/S0002-9440(10)65689-1
  10. Shang R, Lee S, Senavirathne G, Lai EC. microRNAs in action: biogenesis, function and regulation. Nat Rev Genet. 2023;24(12):816–833. doi: 10.1038/s41576-023-00611-y
  11. Chauhan P, Pramodh S, Hussain A, et al. Understanding the role of miRNAs in cervical cancer pathogenesis and therapeutic responses. Front Cell Dev Biol. 2024;12:1397945. doi: 10.3389/fcell.2024.1397945
  12. Robinson M, McCarthy D, Smyth G. edgeR: a Bioconductor package for differential expression analysis of digital gene expression data. Bioinformatics. 2010;26(1):139–40. doi: 10.1093/bioinformatics/btp616
  13. Goedhart J, Luijsterburg MS. VolcaNoseR is a web app for creating, exploring, labeling and sharing volcano plots. Sci Rep. 2020;10(1):20560. doi: 10.1038/s41598-020-76603-3
  14. Xu F, Wang Y, Ling Y, et al. dbDEMC 3.0: Functional Exploration of Differentially Expressed miRNAs in Cancers of Human and Model Organisms. Genomics Proteomics Bioinformatics. 2022;20(3):446–454. doi: 10.1016/j.gpb.2022.04.006
  15. Tang D, Chen M, Huang X, et al. SRplot: A free online platform for data visualization and graphing. PLoS One. 2023;18(11):e0294236. doi: 10.1371/journal.pone.0294236
  16. Kern F, Aparicio-Puerta E, Li Y, et al. miRTargetLink 2.0-interactive miRNA target gene and target pathway networks. Nucleic Acids Res. 2021;49(W1):W409–W16. doi: 10.1093/nar/gkab297
  17. Skoufos G, Kakoulidis P, Tastsoglou S, et al. TarBase-v9.0 extends experimentally supported miRNA-gene interactions to cell-types and virally encoded miRNAs. Nucleic Acids Res. 2024;52(D1):D304–D10. doi: 10.1093/nar/gkad1071
  18. Vasaikar S, Straub P, Wang J, Zhang B. LinkedOmics: analyzing multi-omics data within and across 32 cancer types. Nucleic Acids Res. 2018;46(D1):D956–D63. doi: 10.1093/nar/gkx1090
  19. Zhou Y, Zhou B, Pache L, et al. Metascape provides a biologist-oriented resource for the analysis of systems-level datasets. Nat Commun. 2019;10(1):1523. doi: 10.1038/s41467-019-09234-6
  20. Goldman M, Craft B, Hastie M, et al. Visualizing and interpreting cancer genomics data via the Xena platform. Nat Biotechnol. 2020;38(6):675–678. doi: 10.1038/s41587-020-0546-8
  21. Cruz-De la Rosa M, Jimenez-Wences H, Alarcon-Millan J, et al. miR-218-5p/RUNX2 Axis Positively Regulates Proliferation and Is Associated with Poor Prognosis in Cervical Cancer. Int J Mol Sci. 2022;23(13):6993. doi: 10.3390/ijms23136993
  22. Zhu L, Tu H, Liang Y, Tang D. MiR-218 produces anti-tumor effects on cervical cancer cells in vitro. World J Surg Oncol. 2018;16(1):204. doi: 10.1186/s12957-018-1506-3
  23. Wang P, Zhai G, Bai Y. Values of miR-34a and miR-218 expression in the diagnosis of cervical cancer and the prediction of prognosis. Oncol Lett. 2018;15(3):3580–3585. doi: 10.3892/ol.2018.7791
  24. Kogo R, How C, Chaudary N, et al. The microRNA-218~Survivin axis regulates migration, invasion, and lymph node metastasis in cervical cancer. Oncotarget. 2015;6(2):1090–1100. doi: 10.18632/oncotarget.2836
  25. Yamamoto N, Kinoshita T, Nohata N, et al. Tumor suppressive microRNA-218 inhibits cancer cell migration and invasion by targeting focal adhesion pathways in cervical squamous cell carcinoma. Int J Oncol. 2013;42(5):1523–1532. doi: 10.3892/ijo.2013.1851
  26. Szekerczes T, Galamb A, Varga N, et al. Increased miR-20b Level in High Grade Cervical Intraepithelial Neoplasia. Pathol Oncol Res. 2020;26(4):2633–2640. doi: 10.1007/s12253-020-00852-w
  27. Cheng Y, Geng L, Zhao L, et al. Human papillomavirus E6-regulated microRNA-20b promotes invasion in cervical cancer by targeting tissue inhibitor of metalloproteinase 2. Mol Med Rep. 2017;16(4):5464–5470. doi: 10.3892/mmr.2017.7231
  28. Lajer C, Garnaes E, Friis-Hansen L, et al. The role of miRNAs in human papilloma virus (HPV)-associated cancers: bridging between HPV-related head and neck cancer and cervical cancer. Br J Cancer. 2012;106(9):1526–1534. doi: 10.1038/bjc.2012.109
  29. Wald A, Hoskins E, Wells S, et al. Alteration of microRNA profiles in squamous cell carcinoma of the head and neck cell lines by human papillomavirus. Head Neck. 2011;33(4):504–512. doi: 10.1002/hed.21475
  30. Haga R, Ridley A. Rho GTPases: Regulation and roles in cancer cell biology. Small GTPases. 2016;7(4):207–221. doi: 10.1080/21541248.2016.1232583
  31. Castro-Munoz L, Rocha-Zavaleta L, Lizano M, et al. Alteration of the IFN-Pathway by Human Papillomavirus Proteins: Antiviral Immune Response Evasion Mechanism. Biomedicines. 2022;10(11):2965. doi: 10.3390/biomedicines10112965

Supplementary files

Supplementary Files
Action
1. JATS XML
2. Fig. 1. Next Generation Sequencing results. a. Volcano diagram showing the microRNA expression pattern in СС tumor tissue. The signatures indicate the microRNAs with the most significant expression change in terms of both severity and significance level. b, c. Venn diagrams showing the intersection of the lists of differentially expressed microRNAs in СС according to TCGA and our own data depending on whether the level of microRNAs in tumor tissue is decreased (b) or increased (c).

Download (325KB)
3. Fig. 2. Schematic representation of the algorithm for microRNA target gene search and functional category enrichment. Genes used in the underlying analysis are shown in yellow in the Venn diagram. DE-microRNA — differentially expressed microRNAs.

Download (195KB)
4. Fig. 3: Chord diagram showing the functional categories by which the target genes of the investigated microRNAs are enriched. The left half of the diagram shows the category names, while the right half shows the microRNAs in different colors.

Download (880KB)
5. Fig. 4. Sankey plot showing the most general functional categories of target genes (left part of the plot) corresponding to the microRNAs that regulate them (right part of the plot).

Download (515KB)
6. Fig. 5. Bubble plot of functional categories of target genes corresponding to microRNAs with increased expression in CC tumor tissue. The categories are arranged on the ordinate axis in descending order according to the significance level of the participation of the target genes of the corresponding microRNAs (-log10 <2 on the abscissa axis).

Download (775KB)
7. Fig. 6. Bubble plot of functional categories of target genes corresponding to microRNAs with decreased expression in CC tumor tissue. The categories are arranged on the ordinate axis in descending order according to the significance level of the participation of the target genes of the corresponding microRNAs (–log10 <2 on the abscissa axis).

Download (655KB)
8. Fig. 7. Dependency of the expression level of microRNA-20b (a), microRNA-345 (b) and microRNA-363 (c) on the presence of distant metastases. *p <0,05.

Download (127KB)
9. Fig. 8. Dependency of microRNA-106b expression on tumor malignancy grade. * p <0.05, ** p <0.01.

Download (71KB)
10. Fig. 9. Correlation of microRNA-218-1 expression level with overall survival of patients with CC (a), disease stage (b), and primary tumor size (с). Correlation of microRNA-218-2 expression level with overall survival of patients with CC (d), disease stage (e), and primary tumor size (f). * p <0.05, ** p <0.01, *** p <0.001.

Download (90KB)

Copyright (c) 2024 Eco-Vector



СМИ зарегистрировано Федеральной службой по надзору в сфере связи, информационных технологий и массовых коммуникаций (Роскомнадзор).
Регистрационный номер и дата принятия решения о регистрации СМИ: ПИ № ФС 77 - 86496 от 11.12.2023 г
СМИ зарегистрировано Федеральной службой по надзору в сфере связи, информационных технологий и массовых коммуникаций (Роскомнадзор).
Регистрационный номер и дата принятия решения о регистрации СМИ: ЭЛ № ФС 77 - 80673 от 23.03.2021 г
.