Rondienny Andrade Filgueiras¹, Rosyana de Fátima Vieira de Albuquerque¹, Larissa Silva dos Santos ¹, Alessandra Silva e Silva¹, Cláudia Patrícia Mendes de Araújo¹, Luana Quadros de Souza Leão², Giovana Melo Marques¹, Kamila Pereira de Araújo¹, Jackeline da Silva Luciano¹, Fabiana Ferreira Rosa³, Daniel Lourenço Lira ⁴, Higino Felipe Figueiredo⁴, Mikele Praia de Oliveira⁴, Kátia Luz Torres⁴, Priscila Ferreira de Aquino¹

Introduction: Cervical cancer (CC) is a major cause of cancer-related morbidity and mortality among women, especially in developing countries. In Amazonas, Brazil, CC is the most prevalent malignancy among women, with incidence and mortality rates above national averages. Persistent infection with high-risk human papillomavirus (HR-HPV), especially types 16 and 18, contributes to over 70% of CC cases. These genotypes express oncoproteins E6 and E7, which inactivate tumor suppressors like p53 and Rb, promoting genomic instability and transformation. Despite prevention efforts, biomarkers to predict tumor progression or treatment response remain insufficient, limiting personalized clinical management. This study aimed to characterize the proteomic profile of exfoliated cervical cells from women with advanced-stage CC and HPV-16/18 coinfection in Amazonas, Brazil, and compare it to samples from HPV-negative women with normal cytology (NILM), to identify proteins associated with viral oncogenesis and tumor progression. Materials and Methods: Exfoliated cervical cells from eight women with histologically confirmed advanced CC and HPV-16/18 coinfection (coinfected group) and six HPV-negative women with NILM cytology (women without lesions) were analyzed. Samples were collected using Liqui-PREP® and stored at –80 °C. Proteins were extracted using 0.1% RapiGest™ and mechanical lysis, digested with trypsin, and analyzed via nano-LC-MS/MS (Orbitrap Exploris 480). Protein identification and quantification were performed using PatternLab for Proteomics®, and functional annotation was done using Atlas Proteomics and complementary tools. Results: A total of 492 proteins with spectral count ≥3 were identified, including 186 in the coinfected group (advanced cervical cancer with HPV-16/18) and 451 in the control group (HPV-negative women without lesions), with 145 proteins shared between groups. Fourteen proteins were exclusively identified in the coinfected group, such as PSMB5, PSMB7, proteasome subunit beta type-6, SKP1 (component of the SCF complex), and RAD23A — all related to tumor suppressor degradation and DNA damage response. T-Fold analysis revealed several proteins with statistically significant differential abundance as Peroxiredoxin-2, Apolipoprotein A-I, Apolipoprotein A-II, and inter-alpha-trypsin inhibitor heavy chain H1 were significantly more abundant in the cancer group, suggesting roles in oxidative stress regulation, lipid metabolism, and inflammatory response—reflecting an active and metabolically deregulated tumor microenvironment. In contrast, proteins such as Myeloperoxidase (MPO), Vimentin (VIM), and Immunoglobulin J chain (JCHAIN) were less abundant in the cancer group. These proteins are linked to epithelial integrity, immune surveillance, and antimicrobial defense, indicating a potential loss of these protective functions during tumor progression. Conclusion: Distinct proteomic differences were observed between cervical cells from women with advanced CC and HPV-16/18 coinfection and those from HPV-negative controls. The cancer group exhibited proteins involved in proteostasis disruption, DNA repair, oxidative stress, and immune modulation—key elements of HPV-driven carcinogenesis. These proteins represent potential biomarkers for tumor progression and targets for precision oncology.

Agradecimentos: The authors thanks FAPEAM (POSGRAD Program), CAPES (Funding Code 001), and PROEP/ILMD-FIOCRUZ AMAZÔNIA – LDMAIS for financial support