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Antineoplastic action of sulforaphane on HeLa cells by modulation of signaling pathways and epigenetic pathways
Madhumitha Kedhari Sundaram1, Abdulmajeed G Almutary2, Ahmad Alsulimani3
1School of Life Sciences, Manipal Academy of Higher Education, Dubai, United Arab Emirates.
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 5'CpG 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.
Insights
Sulforaphane, a natural compound, impacts cancer cell pathways by altering gene expression and epigenetic enzyme activity. This research reveals its potential anticancer effects through demethylation and re-expression of tumor suppressor genes in HeLa cells.
Area of Science:
- Epigenetics
- Molecular Biology
- Cancer Research
Background:
- Epigenetic modifications are crucial in altering cellular signaling and molecular pathways.
- These modifications represent significant therapeutic targets in cancer treatment.
- This study investigates the effects of sulforaphane on molecular targets within HeLa cells.
Purpose of the Study:
- To investigate the impact of sulforaphane on gene transcription and epigenetic enzyme activity in HeLa cells.
- To elucidate the molecular mechanisms underlying the potential anticancer effects of sulforaphane.
- To analyze the in-silico interactions and pathway modulations induced by sulforaphane treatment.
Main Methods:
- Quantitative PCR was used to assess the transcriptional levels of various genes.
- Enzyme activity assays (ELISA) measured the function of key epigenetic enzymes.
- Molecular docking and in-silico network analysis were employed to predict interactions and pathway involvement.
Main Results:
- Sulforaphane modulated genes associated with proliferation, metastasis, inflammation, and signal transduction.
- The compound reduced the activity of DNA methyltransferases, histone deacetylases, and histone methyltransferases.
- Demethylation and subsequent re-expression of tumor suppressor genes were observed, correlating with epigenetic enzyme modulation.
Conclusions:
- Sulforaphane treatment leads to significant transcriptional changes in epigenetic regulators and tumorigenesis-related genes.
- The observed demethylation and re-expression of tumor suppressor genes highlight a key mechanism of sulforaphane's anticancer action.
- Findings underscore sulforaphane's potential as a therapeutic agent targeting epigenetic and signaling pathways in cancer.
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