MSc

The study entitled “Anticancer activity of Clitoria ternatea L. through epigenetic regulation” was carried out in the Department of Molecular Biology and Biotechnology, College of Agriculture, Vellayani, Thiruvananthapuram during 2022 - 2023. C. ternatea is a perennial herbaceous plant belonging to the family Fabaceae and it is reported to have anti-oxidant, anti-proliferative, and cytotoxic effects. However, its role in modulation of key epigenetic mechanism namely methylation and the difference in expression of genes associated with methylation has not yet been studied. Hence the objective of this study was to investigate the in vitro anticancer activity of Clitoria ternatea L. and analysing its role on modulation of methylation and differential expression of genes involved in methylation.
Leaves of C. ternatea were dried and extracted with methanol using both cold and soxhlet extraction methods. 640mg and 2510mg of crude extract were obtained from 10g of powdered leaves sample by cold and soxhlet extractions respectively. Cytotoxic effect of the leaf extract was evaluated by MTT assay in human colon cancer cell line (HCT-116), breast cancer cell line (MCF-7), and normal cell line (HEK-293). The extracts showed dose-dependent reduction in cell survival percentage and IC50 values of cold extract were found to be 189.19µg and 176.15µg for colon and breast cancer respectively. The IC50 values of soxhlet extracted methanolic leaf extract were found to be 146.62µg and 148.34µg for colon and breast cancer cell lines respectively. MTT assay on HEK-293 cell line revealed the extracts did not show cytotoxicity up to IC50 values. Soxhlet extracted methanolic leaf extract has higher cytotoxicity than cold extract. Hence soxhlet extract was used for further experiments.
On analysing the global DNA methylation pattern of treated cells with respect to control it was found that there was hypermethylation of 1.20% and 0.76% in colon and breast cancer cell lines respectively. However, there was no change in the methylation pattern of mTOR gene on treated cells. Expression of the methylation specific genes DNMT1, DNMT3a, DNMT3b, MeCP2, and TET2 indicated that hypermethylating genes (DNMT1, DNMT3a, DNMT3b, and MeCP2) were upregulated and the demethylating gene (TET2) was downregulated in treated cells.
The findings suggest that the extract is capable of modulating the methylation profile of cells by regulating the expression of methylation specific genes and C. ternatea can be considered as a good candidate for the development of cancer therapeutics.

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