Sains Malaysiana 54(3)(2025): 839-868

http://doi.org/10.17576/jsm-2025-5403-18

Potentially Dysregulated Cholesterol, Cellular Interaction, Immune, and Collagen in NTCU-Induced Lung Squamous Cell Carcinoma in vivo and LUSC Patients

(Kolesterol Berpotensi Disregulasi, Interaksi Sel, Imun dan Kolagen dalam Karsinoma Sel Skuamosa Paru-paru Aruhan NTCU in vivo dan Pesakit LUSC)

MUHAMMAD ASYAARI ZAKARIA^1,4, AMNANI AMINUDDIN², NOR FADILAH RAJAB³, SITI FATHIAH MASRE^1,* & ENG WEE CHUA²

¹Centre for Toxicology and Health Risk Studies, Faculty of Health Sciences, Universiti Kebangsaan Malaysia, 50300 Kuala Lumpur, Malaysia

²Centre for Drug and Herbal Development, Faculty of Pharmacy, Universiti Kebangsaan Malaysia, 50300 Kuala Lumpur, Malaysia

³Centre for Healthy Ageing and Wellness, Faculty of Health Sciences, Universiti Kebangsaan Malaysia, 50300 Kuala Lumpur, Malaysia

⁴Faculty of Pharmacy and Health Sciences, Universiti Kuala Lumpur-Royal College of Medicine Perak, 30450 Ipoh, Perak, Malaysia

Received: 10 October 2024/Accepted: 26 November 2024

Abstract

Lung squamous cell carcinoma (LUSC) is a deadly cancer, characterized by its complex genetic profiles. Additionally, the molecular mechanisms and etiology underlying LUSC growth are less extensively characterized as compared to adenocarcinoma subtype of lung cancer. Therefore, it is essential to elucidate the molecular mechanisms of LUSC in vivo and by using the human database to understand the disease. A LUSC BALB/c mice model was established using N-nitroso-tris-chloroethylurea (NTCU). After termination of mice, the lung tissues were subjected to RNA sequencing, followed by gene set enrichment analysis (GSEA) to identify the enriched pathways. Subsequently, the pathogenic single nucleotide polymorphism (SNP) was determined and enriched using g:Profiler. The transcriptomic profile of human LUSC patients was obtained and analyzed from The International Cancer Genome Consortium (ICGC). The impact of pathogenic simple somatic mutation (SSM) in human LUSC was determined using the Combined Annotation Dependent Depletion (CADD) score, which was also enriched using g:Profiler. Additionally, the enriched pathway of ‘Treatment-responsive’ was compared with ‘Non-responsive’ LUSC patients’ post-treatment. All pathway analysis was referred to the Reactome database, and an adjusted p-value ≤ 0.05 was considered statistically significant. The top pathway enriched in both mice and human LUSC showed that cholesterol, cellular interaction, immune system, and collagen were significantly affected. Briefly, this study identified important biological pathways that may contribute to LUSC development and hold potential as targets for LUSC therapy in the future.

Keywords: Gene set enrichment analysis (GSEA); lung squamous cell carcinoma (LUSC); RNA sequencing; simple somatic mutation (SSM); single nucleotide polymorphism (SNP)

Abstrak

Karsinoma sel skuamus paru-paru (LUSC) adalah kanser yang boleh membawa maut, dicirikan oleh profil genetiknya yang kompleks. Tambahan pula, mekanisme molekul dan etiologi yang mendasari pertumbuhan LUSC adalah kurang dikaji secara mendalam berbanding subjenis adenokarsinoma kanser paru-paru. Oleh itu, adalah penting untuk mengetahui mekanisme molekul LUSC secara in vivo dan dengan menggunakan pangkalan data manusia untuk memahami penyakit ini. Model tikus BALB/c LUSC telah dibangunkan menggunakan N-nitroso-tris-kloroetilureum (NTCU). Selepas mencit dikorbankan, tisu paru-paru dianalisis dengan penjujukan RNA, diikuti dengan analisis pengayaan set gen (GSEA) untuk mengenal pasti laluan yang diperkaya. Seterusnya, polimorfisme nukleotida tunggal (SNP) yang patogen telah ditentukan dan diperkaya menggunakan g:Profiler. Profil transkriptomik pesakit LUSC manusia diperoleh dan dianalisis daripada Konsortium Genom Kanser Antarabangsa (ICGC). Kesan mutasi somatik ringkas (SSM) yang patogen dalam LUSC manusia ditentukan menggunakan skor Pengurangan Bergantung Anotasi Gabungan (CADD), yang juga diperkaya menggunakan g:Profiler. Tambahan lagi, laluan yang diperkaya bagi pesakit LUSC 'Responsif-rawatan' dibandingkan dengan pesakit 'Tidak responsif' selepas rawatan. Semua analisis laluan dirujuk kepada pangkalan data Reactome dan nilai p terlaras ≤ 0.05 dianggap signifikan secara statistik. Laluan teratas yang diperkaya dalam kedua-dua LUSC model mencit dan manusia mendedahkan penjejasan signifikan kolesterol, interaksi sel, sistem imun dan kolagen. Ringkasnya, kajian ini mengenal pasti laluan biologi penting yang mungkin menyumbang kepada perkembangan LUSC dan berpotensi dijadikan sebagai sasaran terapi LUSC pada masa hadapan.

Kata kunci: Analisis pengayaan set gen (GSEA); karsinoma sel skuamus paru-paru (LUSC); mutasi somatik ringkas (SSM); penjujukan RNA; polimorfisme nukleotida tunggal (SNP)

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*Corresponding author; email: sitifathiah@ukm.edu.my

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