Molecular features associated with progression of stage I-II cervical cancer

Background. The treatment efficacy of stage I-II cervical cancer (CC) remains suboptimal due to the risk of recurrence and metastasis, which complicates the selection of optimal treatment strategies and necessitates the search for prognostic markers.

This study aimed to identify molecular characteristics and markers associated with progressive stage I-II CC.

Material and Methods. Clinical and morphological analyses and whole-transcriptome sequencing were performed, and publicly available data from the Cancer Genome Atlas (TCGA) database were used to validate the sequencing results.

Results. Patients with progressive disease demonstrated activation of oncogenic (TNF-α via NF-κB, PI3K/AKT/mTOR, MTORC1, and KRAS), metabolic (oxidative phosphorylation, glycolysis, and hypoxia), immune (TGF-β, responses to IFNγ and IFNα, and IL2/STAT5), and G2/M cell cycle (through regulation of the E2F and MYC transcription factor targets) signaling pathways. the ITGA6 gene expression level was higher in tumors from patients with cervical cancer progression than in patients without cancer progression.

Conclusion. The progression of stage I-II cervical cancer is driven by a complex interplay of proliferative, metabolic, and immunosuppressive factors, with ITGA6 identified as a potential marker.

1. Siegel R.L., Kratzer T.B., Giaquinto A.N., Sung H., Jemal A. Cancer statistics, 2025. CA Cancer J Clin. 2025; 75(1): 10–45. doi:10.3322/caac.21871.

2. Arbyn M., Weiderpass E., Bruni L., de Sanjosé S., Saraiya M., Ferlay J., Bray F. Estimates of incidence and mortality of cervical cancer in 2018: a worldwide analysis. Lancet Glob Health. 2020; 8(2): e191–e203. doi: 10.1016/s2214-109x(19)30482-6.

3. Flint M., Yi F., Saleh M., Liu X., Blank S.V., Liu Y. Cervical cancer in women under age 40: A cohort study of clinicopathological characteristics and fertility-sparing surgery outcomes. Gynecol Oncol Rep. 2025; 58: 101710. doi: 10.1016/j.gore.2025.101710.

4. Caruso G., Wagar M.K., Hsu H.C., Hoegl J., Rey Valzacchi G.M., Fernandes A., Cucinella G., Sahin Aker S., Jayraj A.S., Mauro J., Pareja R., Ramirez P.T. Cervical cancer: a new era. Int J Gynecol Cancer. 2024; 34(12): 1946–70. doi: 10.1136/ijgc-2024-005579.

5. Kravets O.A., Romanova E.A., Gorbunova V.A. Clinical results of radiation and chemoradiation therapy of locally advanced cervix cancer. Russian Journal of Oncology. 2020; 25(3): 92–102. (in Russian). doi: 10.17816/1028-9984-2020-25-3-92-102. EDN: THVKUP.

6. Reyes Santiago D.К., Khadzhimba A.S., Sobolev I.V., Ilyin A.A., Maksimov S.Ya. Outcomes of combination therapy for stage IIA–IIIB cervical cancer. Tumors of Female Reproductive System. 2018; 14(4): 50–55. (in Russian). doi: 10.17650/1994-4098-2018-14-4-50-55. EDN: YXHEAH.

7. Ecker S., Kirisits C., Schmid M., De Leeuw A., Seppenwoolde Y., Knoth J., Trnkova P., Heilemann G., Sturdza A., Kirchheiner K., Spampinato S., Serban M., Jürgenliemk-Schulz I., Chopra S., Nout R., Tanderup K., Pötter R., Eder-Nesvacil N. Tools for large-scale data analytics of an international multi-center study in radiation oncology for cervical cancer. Radiother Oncol. 2023; 182: 109524. doi: 10.1016/j.radonc.2023.109524.

8. Liu X., Meng Q., Wang W., Zhou Z., Zhang F., Hu K. Predictors of Distant Metastasis in Patients with Cervical Cancer Treated with Definitive Radiotherapy. J Cancer. 2019; 10(17): 3967–74. doi: 10.7150/jca.31538.

9. Wu C., Lv X., Wang F., Xu Q., Lou H., Zhang X. Predictors of distant metastasis or local recurrent after radiotherapy in patients with cervical cancer. BMC Cancer. 2025; 25(1): 364. doi: 10.1186/s12885-024-13282-7.

10. Chen X., Dong X., Li H., Wu T., Liu H., Wu J., Ge W., Hao L., Zhang Z. RNA-binding proteins signature is a favorable biomarker of prognosis, immunotherapy and chemotherapy response for cervical cancer. Cancer Cell Int. 2024; 24(1): 80. doi: 10.1186/s12935-024-03257-w.

11. Li J., Hou F., Teng Z., Xia W., Peng J. LncRNA HOXC-AS3 accelerates malignant proliferation of cervical cancer cells via stabilizing KDM5B. J Cancer Res Clin Oncol. 2024; 150(6): 294. doi: 10.1007/ s00432-024-05799-y.

12. Li Y., Hong Y., Shen H., Zhou J., Cesar D., Eleutério J.Jr., Matsuura M., Liu Y., Luo C., Li Q. FXR activation suppresses NF-κB signaling, proliferation and migration in cervical cancer cells. Transl Cancer Res. 2025; 14(4): 2440–56. doi: 10.21037/tcr-2025-522.

13. Hu D., Zhang Z., Zhang Y., Huang K., Li X. Identification of immune related molecular subtypes and prognosis model for predicting prognosis, drug resistance in cervical squamous cell carcinoma. Front Genet. 2023; 14: 1137995. doi: 10.3389/fgene.2023.1137995.

14. Feng S., Wang Z., Zhang H., Hou B., Xu Y., Hao S., Lu Y. Identification of prognostic biomarkers for cervical cancer based on programmed cell death-related genes and assessment of their immune profile and response to drug therapy. J Gene Med. 2024; 26(1): e3643. doi:10.1002/jgm.3643.

15. Sun K., Huang C., Li J.Z., Luo Z.X. Identification of a necroptosisrelated prognostic gene signature associated with tumor immune microenvironment in cervical carcinoma and experimental verification. World J Surg Oncol. 2022; 20(1): 342. doi:10.1186/s12957-022-02802-z.

16. Xing X., Tian Y., Jin X. Immune infiltration and a necroptosisrelated gene signature for predicting the prognosis of patients with cervical cancer. Front Genet. 2023; 13. doi: 10.3389/fgene.2022.1061107.

17. Fleischmann M., Chatzikonstantinou G., Fokas E., Wichmann J., Christiansen H., Strebhardt K., Rödel C., Tselis N., Rödel F. Molecular Markers to Predict Prognosis and Treatment Response in Uterine Cervical Cancer. Cancers. 2021; 13(22): 5748.

18. WHO Classification of Tumours of Female Reproductive Organs. Ed. R.J. Kurman, M.L. Carcangiu, C.S. Herrington, R.H. Young. 4th Edition. 2014. Vol. 6. ISBN: 978-92-832-2435-8.

19. Khokhlova S.V., Kravets O.A., Morkhov K.Yu., Nechushkina V.M., Saevets V.V., Tyulyandina A.S., Ulrich E.A., Usmanova L.Sh. Practical recommendations for the drug treatment of cervical cancer. Malignant Tumors. 2022; 12(3S2-1): 240–59. (in Russian). doi: 10.18027/2224-5057-2022-12-3s2-240-259. EDN: EETIUT.

20. Ewels P., Magnusson M., Lundin S., Käller M. MultiQC: summarize analysis results for multiple tools and samples in a single report. Bioinformatics.2016; 32(19): 3047–48. doi: 10.1093/bioinformatics/btw354.

21. Dobin A., Davis C.A., Schlesinger F., Drenkow J., Zaleski C., Jha S., Batut P., Chaisson M., Gingeras T.R. STAR: ultrafast universal RNA-seq aligner. Bioinformatics. 2013; 29(1): 15–21. doi: 10.1093/bioinformatics/bts635.

22. Putri G.H., Anders S., Pyl P.T., Pimanda J.E., Zanini F. Analysing high-throughput sequencing data in Python with HTSeq 2.0. Bioinformatics. 2022; 38(10): 2943–45. doi: 10.1093/bioinformatics/btac166.

23. Love M.I., Huber W., Anders S. Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2. Genome Biol. 2014; 15(12): 550. doi: 10.1186/s13059-014-0550-8.

24. Korotkevich G., Sukhov V., Budin N., Shpak B., Artyomov M.N., Sergushichev A. Fast gene set enrichment analysis. bioRxiv.2021. https://doi.org/10.1101/060012.

25. Zhen Y, Pavez M., Li X. The role of Pcdh10 in neurological disease and cancer. J Cancer Res Clin Oncol. 2023; 149(10): 8153–64. doi: 10.1007/s00432-023-04743-w.

26. Hu D.G., Marri S., Hulin J-A., McKinnon R.A., Mackenzie P.I., Meech R. The Somatic Mutation Landscape of UDP-Glycosyltransferase (UGT) Genes in Human Cancers. Cancers. 2022; 14(22): 5708. doi: 10.3390/cancers14225708.

27. Zhu Y., Wu Y., Yang L., Dou X., Jiang J., Wang L. Long non-coding RNAactivated by transforming growth factor-β promotes proliferation and invasion of cervical cancer cells by regulating the miR-144/ITGA6 axis. Exp Physiol. 2019; 104(6): 837–44. doi: 10.1113/ep087656.

28. Khademi R., Malekzadeh H., Bahrami S., Saki N., Khademi R., Villa-Diaz L.G. Regulation and Functions of α6-Integrin (CD49f) in Cancer Biology. Cancers. 2023; 15(13): 3466. doi: 10.3390/cancers15133466.

29. Du J., Luo H., Ye S., Zhang H., Zheng Z., Liu K. Unraveling IFI44L’s biofunction in human disease. Front Oncol. 2024; 14: 1436576. doi: 10.3389/fonc.2024.1436576.

30. Fu T., Liu J.X., Xie J., Gao Z., Yang Z. LAMC2 as a prognostic biomarker in human cancer: a systematic review and meta-analysis. BMJ Open. 2022; 12(11): e063682. doi: 10.1136/bmjopen-2022-063682.

31. Gong T.T., Liu F.H., Xiao Q., Li Y.Z., Wei Y.F., Xu H.L, Cao F., Sun M.L, Jiang F.L., Tao T., Ma Q.P., Qin X., Song Y., Gao S., Wu L., Zhao Y.H., Huang D.H., Wu Q.J. SH3RF2 contributes to cisplatin resistance in ovarian cancer cells by promoting RBPMS degradation. Commun Biol. 2024; 7(1): 67. doi: 10.1038/s42003-023-05721-1.

32. Lin X., Yoshikawa N., Liu W., Matsukawa T., Nakamura K., Yoshihara M., Koya Y., Sugiyama M., Tamauchi S., Ikeda Y., Yokoi A., Shimizu Y., Kajiyama H.. DDIT4 Facilitates Lymph Node Metastasis via the Activation of NF-κB Pathway and Epithelial-Mesenchymal Transition. Reprod Sci. 2023; 30(9): 2829–41. doi: 10.1007/s43032-023-01230-y.

33. Hakim R.U., Amin T., U.l. Islam S.M.B. Advances and Challenges in Cervical Cancer: From Molecular Mechanisms and Global Epidemiology to Innovative Therapies and Prevention Strategies. Cancer Control. 2025; 32: 10732748251336415. doi: 10.1177/10732748251336415.

34. Zhou Y., Rassy E., Coutte A., Achkar S., Espenel S., Genestie C., Pautier P., Morice P., Gouy S., Chargari C. Current Standards in the Management of Early and Locally Advanced Cervical Cancer: Update on the Benefit of Neoadjuvant/Adjuvant Strategies. Cancers (Basel). 2022; 14(10): 2449. doi: 10.3390/cancers14102449.

35. Yang C., Zhang Z.C., Liu T.B., Xu Y., Xia B.R., Lou G. E2F1/2/7/8 as independent indicators of survival in patients with cervical squamous cell carcinoma. Cancer Cell Int. 2020; 20: 500. doi: 10.1186/s12935-020-01594-0.

36. Li H.L., Dong L.L., Jin M.J., Li Q.Y., Wang X., Jia M.Q., Song J., Zhang S.Y., Yuan S. A Review of the Regulatory Mechanisms of N-Myc on Cell Cycle. Molecules. 2023; 28(3): 1141.

37. Gao Y., Qiao X., Liu Z., Zhang W. The role of E2F2 in cancer progression and its value as a therapeutic target. Front Immunol. 2024; 15: 1397303. doi: 10.3389/fimmu.2024.1397303.

38. Liu Y., Wang Y., Tan S., Shi X., Wen J., Chen D., Zhao Y., Pan W., Jia Z., Lu C., Lou G. Characterization of G2/M checkpoint classifier for personalized treatment in uterine corpus endometrial carcinoma. Cancer Cell Intern. 2025; 25(1): 34. doi: 10.1186/s12935-025-03667-4.

39. Lin X., Fang Y., Mi X., Fu J., Chen S., Wu M., Jin N. Asiatic acid inhibits cervical cancer cell proliferation and migration via PI3K/AKT/ mTOR signaling pathway. Heliyon. 2024; 10(13): e34047. doi: 10.1016/j.heliyon.2024.e34047.

40. Ueda H., Ishiguro T., Mori Y., Yamawaki K., Okamoto K., Enomoto T., Yoshihara K. Glycolysis-mTORC1 crosstalk drives proliferation of patientderived endometrial cancer spheroid cells with ALDH activity. Cell Death Disc. 2024; 10(1): 435. doi: 10.1038/s41420-024-02204-y.

41. Datta A., West C., O’Connor J.P.B., Choudhury A., Hoskin P. Impact of hypoxia on cervical cancer outcomes. Int J Gynecol Cancer. 2021; 31(11): 1459–70. doi: 10.1136/ijgc-2021-002806.

42. Abdel-Wahab A.F., Mahmoud W., Al-Harizy R.M. Targeting glucose metabolism to suppress cancer progression: prospective of anti-glycolytic cancer therapy. Pharmacol Res. 2019; 150: 104511. doi: 10.1016/j.phrs.2019.104511.

43. Sheng X., Wang M.-M., Zhang G.-D., Su Y., Fang H.-B., Yu Z.-H., Su Z. Dual inhibition of oxidative phosphorylation and glycolysis to enhance cancer therapy. Bioorg Chem. 2024; 147: 107325. doi: 10.1016/j.bioorg.2024.107325.

44. Niu N., Ye J., Hu Z., Zhang J., Wang Y. Regulative Roles of Metabolic Plasticity Caused by Mitochondrial Oxidative Phosphorylation and Glycolysis on the Initiation and Progression of Tumorigenesis. Int J Mol Sci. 2023; 24(8): 7076. doi: 10.3390/ijms24087076.

45. Shi X., Yang J., Deng S., Xu H., Wu D., Zeng Q., Wang S., Hu T, Wu F., Zhou H. TGF-β signaling in the tumor metabolic microenvironment and targeted therapies. J Hematol Oncol. 2022; 15(1): 135. doi: 10.1186/s13045-022-01349-6.

46. Hollern D.P., Swiatnicki M.R., Andrechek E.R. Histological subtypes of mouse mammary tumors reveal conserved relationships to human cancers. PLOS Genetics. 2018; 14(1): e1007135. doi: 10.1371/journal.pgen.1007135.

47. Yin S., Cui H., Qin S., Yu S. Manipulating TGF-β signaling to optimize immunotherapy for cervical cancer. Biomed Pharmacother. 2023; 166: 115355. doi: 10.1016/j.biopha.2023.115355.

48. Birrer M.J., Fujiwara K., Oaknin A., Randall L., Ojalvo L.S., Valencia C., Ray-Coquard I. The Changing Landscape of Systemic Treatment for Cervical Cancer: Rationale for Inhibition of the TGF-β and PD-L1 Pathways. Front Oncol. 2022; 12: 814169. doi: 10.3389/fonc.2022.814169.

49. Baba A.B., Rah B., Bhat G.R., Mushtaq I., Parveen S., Hassan R., Hameed Zargar M., Afroze D. Transforming Growth Factor-Beta (TGF-β) Signaling in Cancer-A Betrayal Within. Front Pharmacol. 2022; 13: 791272. doi: 10.3389/fphar.2022.791272.

50. Rangel-Corona R., Corona-Ortega T., Soto-Cruz I., López-Labra A., Pablo-Arcos T., Torres-Guarneros C.F., Weiss-Steider B. Evidence that cervical cancer cells secrete IL-2, which becomes an autocrine growth factor. Cytokine. 2010; 50(3): 273–77. doi:10.1016/j.cyto.2010.02.013.

51. Tilborghs S., Corthouts J., Verhoeven Y., Arias D., Rolfo C., Trinh X.B., van Dam P.A. The role of Nuclear Factor-kappa B signaling in human cervical cancer. Crit Rev Oncol Hematol. 2017; 120: 141–50. doi: 10.1016/j.critrevonc.2017.11.001.

52. Sahin K., Tuzcu M., Basak N., Caglayan B., Kilic U., Sahin F., Kucuk O. Sensitization of Cervical Cancer Cells to Cisplatin by Genistein: The Role of NFκB and Akt/mTOR Signaling Pathways. J Oncol. 2012; 2012: 461562. doi: 10.1155/2012/461562.