Browsing by Author "Sindura, K P"

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    Epigenetic modulation mediated anticancer activity of Pogostemon quadrifolius (Benth)
    (Department of Molecular Biology and Biotechnology, College of Agriculture,Vellayani, 2024-07-31) Keerthana, J; Sindura, K P
    The study entitled ―Epigenetic modulation mediated anticancer activity of Pogostemon quadrifolius (Benth.)‖ was conducted in the Department of Molecular Biology and Biotechnology, College of Agriculture, Vellayani, Thiruvananthapuram from 2023- 2024. Pogostemon quadrifolius (Benth.) (Malayalam name: Naithumba) is a medicinal shrub in the family Lamiaceae and it is reported to have antibacterial, antifungal, antioxidant, and antiproliferative properties. Previous studies from our lab have reported the cytotoxic activity of P. quadrifolius (Benth.) against lung and breast cancer cell lines. However its mechanism of cytotoxic action is not yet thoroughly investigated. Hence the objective was in vitro analysis of epigenetic regulation mediated anticancer activity of Pogostemon quadrifolius (Benth.) and differential expression analysis of genes involved in methylation and acetylation. P. quadrifolius (Benth.) leaves were dried, powdered, and then extracted using methanol by cold extraction method. 403mg of crude extract was obtained from 3g of powdered leaf sample. The human breast cancer cell line (MDA-MB -231), human lung cancer cell line (A549), and normal cell line (HEK-293) were cultured and treated with varying concentrations of methanolic leaf extract and its cytotoxicity on cell lines were determined by MTT assay. The extract showed a dose-dependent reduction in cell viability percentage and the half-maximum inhibitory concentration or IC50 values were found to be 76.22µg/ml and 52.35µg/ml for breast and lung cancer respectively, while it was non-toxic towards normal cells at the obtained IC50 concentrations. Global DNA methylation pattern of P. quadrifolius (Benth.) methanolic leaf extract (PQML) treated lung cancer cells was evaluated and it was found that the relative level of methylation of untreated and treated cells was about 2.54% and 5.76% respectively. The effect of leaf extract on the promoter hypermethylation of the p15, RARβ2, and RASSF1A genes in the breast and lung cancer cell lines was monitored by methylation specific PCR. PQML demethylates the p15 promoter in A549 cell line and RARβ2 promoter in MDA-MB-231 cell line. The RASSF1A gene 89 did not show any change in methylation status on treatment with PQML in both MDA-MB-231 and A549 cell lines. Expression analysis of methylation specific genes showed downregulation of methylation specific genes (DNMT1, DNMT3a, DNMT3b, and MeCP2) and upregulation of the demethylation gene (TET2) in PQML treated breast and lung cancer cells. Expression analysis of histone acetylation/ deacetylation specific genes showed downregulation of acetylation specific (CBP, p300) and deacetylation specific genes (HDAC1, HDAC3) in both cell lines treated with PQML. Our results suggest that the methanolic leaf extract of P. quadrifolius (Benth.) can alter the methylation profile of lung cancer cells as well as regulate the expression levels of genes specific to methylation and acetylation in lung and breast cancer cell lines. It is also found to change the methylation pattern in tumor suppressor genes namely p15 and RARβ2 genes. Hence, P. quadrifolius (Benth.) is proved to have an epigenetic mode of cytotoxic effect on lung and breast cancer cell lines.
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    Establishment of in vitro root cultures of sarsaparilla (Hemidesmus indicus L) R.Br.
    (Department of Plant Biotechnology, College of Agriculture, Vellayani, 2014) Sindura, K P; Satheeshkumar, K