Browsing by Author "Mahek Sharan"

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Demethylation of CADM1 and SOCS1 using capsaicin in cervical cancer cell line
(Springer Science and Business Media Deutschland GmbH, 2023) Mahek Sharan; Meenakshi Jha; Rishima Chandel; Saima Syeda; Runjhun Mathur; Niraj Kumar Jha; Saurabh Kumar Jha; Harsh Goel; Anju Shrivastava; Sushma Chauhan; Sudheer Pamidimarri; Abhimanyu Kumar Jha
Cervical cancer is one of the leading causes of women’s mortality in developing countries. The prevalence of cervical cancer is higher in developing countries like India and continents like Africa. Hyper-methylation of tumor suppressor genes through human papillomavirus (HPV) infection is known to be one of the major causes of cervical cancer. The promoter hypermethylation of the cell adhesion molecule 1 (CADM1) and suppressor of cytokine signalling (SOCS1) genes due to DNMT1 overexpression leads to their epigenetic silencing followed by gene repression causing cervical cancer. In silico study on the inhibition effect of capsaicin on DNMT1 was simulated by different servers. The binding energy was observed to be −7.8 kcal/mol. In vitro studies on the effect of capsaicin on aberrant methylation of CADM1 and SOCS1 were performed on the adenocarcinoma cervical cancer cell line, HeLa. The IC50 of capsaicin was observed to be 160 μM through crystal violet assay. DNA methylation of the CADM1 and SOCS1 was analyzed by methylation-specific PCR along with their reversal using capsaicin (20 μM) by treating the cells for 72 h and 6 days. In silico results suggested that capsaicin has an inhibitory effect on DNMT1, which regulates DNA methylation leading to the hypermethylation of CADM1 and SOCS1 genes. The in vitro studies suggested that hypermethylation leads to the inhibition of CADM1 and SOCS1 expression, which could be reversed using capsaicin with visible changes in methylation-specific and unmethylation-specific bands in MS-PCR, respectively. The present study shows the reversal of methylation of CADM1 and SOCS1 after 72 h which showed a further increase in case of 6 days of treatment using 20 μM capsaicin, which makes capsaicin a potent candidate for causing demethylation of CADM1 and SOCS1 genes that may lead to the reactivation of the downregulated gene. © 2022, The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.
Reversal of promoter hypermethylation of CADM1 and SOCS1 by leaf extract of Datura metel in cervical cancer cells
(ScienceIn Publishing, 2024) Mahek Sharan; Niraj Kumar Jha; Saurabh Kumar Jha; Abhimanyu Kumar Jha
Cervical cancer ranks as the second most fatal cancer among women in developing countries, trailing only behind breast cancer. Human Papillomavirus (HPV) infection, among various contributors, stands as a primary cause of cervical cancer. Overexpression of DNA methyltransferase 1 (DNMT1) leads to hypermethylation of Cell Adhesion Molecule 1 (CADM1) and Suppressor of Cytokine Signaling 1 (SOCS1), consequently silencing these tumor suppressor genes (TSGs) epigenetically. In this study, we explored the reversal of aberrant methylation in the squamous cervical cancer cells, SiHa using a concentration of 5 µg/ml of Datura metel ethanol-chloroform (E: C) leaf extract. Treating the cells with the extract for 72 hours and consecutively for six days, the DNA fragmentation study for cell apoptosis was performed. The methylation-specific PCR analyzed the DNA aberrant methylation patterns of these TSGs and their subsequent reversal and fragmentation. The results suggest that the Datura metel leaf extract (E: C) was able to cause DNA fragmentation and also effectively reversed the promoter hypermethylation, leading to the reactivation of CADM1 and SOCS1. This was evidenced by the reduction in intensity and visual sharpness of the methylation-specific band and the unmethylation-specific band displayed an increase in width and enhanced luminosity in MS-PCR for both genes. This study marks one of the initial global reports showcasing the potential of E: C leaf extract from Datura metel in reversing hypermethylation and reactivating CADM1 and SOCS1 genes in cervical cancer cells. Further exploration into the phytochemicals of D. metel leaves that demethylate CADM1 and SOCS1 could unveil a promising candidate for reactivating suppressed genes in cervical cancer. © ScienceIn Publishing.