ORIGINAL ARTICLE   

Minerva Medica 2021 December;112(6):792-803

DOI: 10.23736/S0026-4806.21.07656-4

Copyright © 2021 EDIZIONI MINERVA MEDICA

lingua: Inglese

Antineoplastic action of sulforaphane on HeLa cells by modulation of signaling pathways and epigenetic pathways

Madhumitha KEDHARI SUNDARAM ¹, Abdulmajeed G. ALMUTARY ², Ahmad ALSULIMANI ³, Syed REHAN AHMAD ⁴, Pallavi SOMVANSHI ^{5, 6}, Tulika BHARDWAJ ⁵, Rinaldo PELLICANO ⁷, Sharmila FAGOONEE ⁸, Arif HUSSAIN ¹ ✉, Shafiul HAQUE ^{9, 10}

¹ School of Life Sciences, Manipal Academy of Higher Education, Dubai, United Arab Emirates; ² College of Applied Medical Sciences, Department of Medical Biotechnology, Qassim University, Buraydah, Saudi Arabia; ³ College of Applied Medical Sciences, Department of Medical Laboratory Technology, Jazan University, Jazan, Saudi Arabia; ⁴ Hiralal Mazumdar Memorial College of Women, Kolkata, India; ⁵ School of Computational & Integrative Sciences (SC&IS), Jawaharlal Nehru University, New Delhi, India; ⁶ Special Centre of Systems Medicine (SCSM), Jawaharlal Nehru University, New Delhi, India; ⁷ Unit of Gastroenterology, Molinette Hospital, Turin, Italy; ⁸ Institute of Biostructure and Bioimaging (CNR), Molecular Biotechnology Center, Turin, Italy; ⁹ Research and Scientific Studies Unit, College of Nursing & Allied Health Sciences, Jazan University, Jazan, Saudi Arabia; ¹⁰ Bursa Uludağ University Faculty of Medicine, Görükle Campus, Nilüfer, Bursa, Turkey

BACKGROUND: Epigenetic modifications alter signaling and molecular pathways; moreover, they are an important therapeutic target. This study examined the effect of sulforaphane on molecular targets in HeLa cells.
METHODS: Quantitative PCR of various molecular targets was performed. Activity of epigenetic enzymes was measured by ELISA and molecular docking analysis was conducted. Promoter methylation of some tumor suppressor genes was quantified using PCR based methylation array. In-silico protein-protein interaction network analysis was performed to understand the effect of transcriptional changes.
RESULTS: Quantitative PCR demonstrated the transcriptional modulation of genes involved in proliferation, metastasis, inflammation, signal transduction pathways and chromatin modifiers. Sulforaphane reduced the enzymatic activity of DNA methyl transferases, histone deacetylases and histone methyltransferases. Molecular docking results suggest that sulforaphane competitively inhibited several DNA methyl transferases and histone deacetylases. Promoter 5ʹCpG methylation levels of selected tumor suppressor genes was found to be reduced which correlated with their transcriptional increase as well modulation of epigenetic enzymes. Further, protein-protein interaction network analysis discerned the participation of genes towards cancer pathways. Functional enrichment and pathway-based analysis represented the modulation of epigenetic and signaling pathways on sulforaphane treatment.
CONCLUSIONS: The modulation in transcriptional status of epigenetic regulators, genes involved in tumorigenesis resulting in tumor suppressor genes demethylation and re-expression underscores the mechanism behind the anticancer effect of sulforaphane on HeLa cells.

KEY WORDS: Epigenomics; Molecular docking simulation; Protein interaction maps; Sulforaphane; Genes, tumor suppressor