Protein Profiling in Human Papillomavirus-Associated Cervical Carcinogenesis: Cornulin as a Biomarker for Disease Progression

Gaayathri Kumarasamy; Mohd Nazri Ismail; Sharifah Emilia Tuan Sharif; Christopher Desire; Parul Mittal; Peter Hoffmann; Gurjeet Kaur

doi:10.3390/cimb45040235

Protein Profiling in Human Papillomavirus-Associated Cervical Carcinogenesis: Cornulin as a Biomarker for Disease Progression

Curr Issues Mol Biol. 2023 Apr 20;45(4):3603-3627. doi: 10.3390/cimb45040235.

Authors

Gaayathri Kumarasamy¹, Mohd Nazri Ismail^{1

2}, Sharifah Emilia Tuan Sharif³, Christopher Desire⁴, Parul Mittal⁴, Peter Hoffmann⁴, Gurjeet Kaur¹

Affiliations

¹ Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, Minden 11800, Pulau Pinang, Malaysia.
² Analytical Biochemistry Research Centre (ABrC), Universiti Sains Malaysia, Bayan Lepas 11900, Pulau Pinang, Malaysia.
³ Department of Pathology, School of Medical Sciences, Universiti Sains Malaysia, Kubang Kerian 16150, Kelantan, Malaysia.
⁴ Clinical Health Sciences, University of South Australia, City West Campus, Adelaide, SA 5000, Australia.

Abstract

Nearly 90% of cervical cancers are linked to human papillomavirus (HPV). Uncovering the protein signatures in each histological phase of cervical oncogenesis provides a path to biomarker discovery. The proteomes extracted from formalin-fixed paraffin-embedded tissues of the normal cervix, HPV16/18-associated squamous intraepithelial lesion (SIL), and squamous cell carcinoma (SCC) were compared using liquid chromatography-mass spectrometry (LC-MS). A total of 3597 proteins were identified, with 589, 550, and 1570 proteins unique to the normal cervix, SIL, and SCC groups, respectively, while 332 proteins overlapped between the three groups. In the transition from normal cervix to SIL, all 39 differentially expressed proteins were downregulated, while all 51 proteins discovered were upregulated in SIL to SCC. The binding process was the top molecular function, while chromatin silencing in the SIL vs. normal group, and nucleosome assembly in SCC vs. SIL groups was the top biological process. The PI3 kinase pathway appears crucial in initiating neoplastic transformation, while viral carcinogenesis and necroptosis are important for cell proliferation, migration, and metastasis in cervical cancer development. Annexin A2 and cornulin were selected for validation based on LC-MS results. The former was downregulated in the SIL vs. normal cervix and upregulated in the progression from SIL to SCC. In contrast, cornulin exhibited the highest expression in the normal cervix and lowest in SCC. Although other proteins, such as histones, collagen, and vimentin, were differentially expressed, their ubiquitous expression in most cells precluded further analysis. Immunohistochemical analysis of tissue microarrays found no significant difference in Annexin A2 expression between the groups. Conversely, cornulin exhibited the strongest expression in the normal cervix and lowest in SCC, supporting its role as a tumor suppressor and potential biomarker for disease progression.

Keywords: biomarker; cervical cancer; human papillomavirus; oncogenesis; proteomics.

Grants and funding

FRGS/1/2019/SKK13/USM/01/1/Ministry of Higher Education