Phyllanthus debilis methanolic extract reduces the viability of human colorectal adenocarcinoma (HT-29) cells and increases LINE-1 and Alu DNA methylation

Siti Nur Dalila Mohd Zain, . and Wan Adnan Wan Omar, . Phyllanthus debilis methanolic extract reduces the viability of human colorectal adenocarcinoma (HT-29) cells and increases LINE-1 and Alu DNA methylation. pp. 25-36. ISSN 1511-3701

Phyllanthus debilis was shown to have a strong anti-proliferative effect on cancer cells with less effect in normal cells. However its mechanism on the epigenetic mechanism at repeat sequences is unknown. This study was carried out to determine the effect of P. debilis extract on long interspersed nuclear element-1 (LINE-1) and Alu DNA methylation. The anti-proliferative effect of P. debilis methanolic extract on human colorectal adenocarcinoma (HT-29) at 24 hours was done using trypan blue assay. LINE-1 and Alu methylation measurement on the HT-29 cell line was done after 72 hours of treatment using Pyrosequencing. The effect of P. debilis methanolic extract at 24 hours on the viability of HT-29 cells was dose-dependent with the half-maximal inhibitory concentration (IC‚‚) concentration of 0.1 mg/mL. Treatment with P. debilis methanolic extract showed significantly higher Alu DNA methylation when compared with the untreated HT-29 cells (37.0 2.5 vs 32.3 4.3 p0.05). Similarly treatment with 5-aza-2-deoxycytidine also significantly increased the Alu DNA methylation compared with the untreated HT-29 cells (46.0 2.3 vs 37.0 2.5 p0.05). For LINE-1 there was a significant increase of LINE-1 methylation when treated with P. debilis extract (80.3 1.3 vs 76.3 2.1 p0.05) and with 5-aza-2-deoxycytidine (81.8 4.3 vs 76.3 2.1 p0.05) when compared with untreated cells. In conclusion treatment of P. debilis methanolic extract on HT-29 cell line reduces the viability of HT-29 cells and increases the methylation of Alu and LINE 1. Similar changes in methylation were also seen in the 5-aza treatment. These epigenetic changes by P. debilis methanolic extract may contribute to its anti-cancer properties.

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Abstract

Phyllanthus debilis was shown to have a strong anti-proliferative effect on cancer cells with less effect in normal cells. However its mechanism on the epigenetic mechanism at repeat sequences is unknown. This study was carried out to determine the effect of P. debilis extract on long interspersed nuclear element-1 (LINE-1) and Alu DNA methylation. The anti-proliferative effect of P. debilis methanolic extract on human colorectal adenocarcinoma (HT-29) at 24 hours was done using trypan blue assay. LINE-1 and Alu methylation measurement on the HT-29 cell line was done after 72 hours of treatment using Pyrosequencing. The effect of P. debilis methanolic extract at 24 hours on the viability of HT-29 cells was dose-dependent with the half-maximal inhibitory concentration (IC‚‚) concentration of 0.1 mg/mL. Treatment with P. debilis methanolic extract showed significantly higher Alu DNA methylation when compared with the untreated HT-29 cells (37.0 2.5 vs 32.3 4.3 p0.05). Similarly treatment with 5-aza-2-deoxycytidine also significantly increased the Alu DNA methylation compared with the untreated HT-29 cells (46.0 2.3 vs 37.0 2.5 p0.05). For LINE-1 there was a significant increase of LINE-1 methylation when treated with P. debilis extract (80.3 1.3 vs 76.3 2.1 p0.05) and with 5-aza-2-deoxycytidine (81.8 4.3 vs 76.3 2.1 p0.05) when compared with untreated cells. In conclusion treatment of P. debilis methanolic extract on HT-29 cell line reduces the viability of HT-29 cells and increases the methylation of Alu and LINE 1. Similar changes in methylation were also seen in the 5-aza treatment. These epigenetic changes by P. debilis methanolic extract may contribute to its anti-cancer properties.

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Item Type:	Article
AGROVOC Term:	Phyllanthus
AGROVOC Term:	Methanol
AGROVOC Term:	Extracts
AGROVOC Term:	Medicinal herbs
AGROVOC Term:	Cell culture
AGROVOC Term:	Methylation
AGROVOC Term:	Viability
AGROVOC Term:	Cancer (disease)
AGROVOC Term:	Disease treatment
Depositing User:	Mr. AFANDI ABDUL MALEK
Last Modified:	24 Apr 2025 00:55
URI:	http://webagris.upm.edu.my/id/eprint/10354

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