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ORIGINAL ARTICLE
Year : 2020  |  Volume : 16  |  Issue : 67  |  Page : 57-60

The effect of Phyllanthus debilis methanolic extract on DNA methylation of TAC1 gene in colorectal cancer cell line


Integrative Medicine Cluster, Advanced Medical and Dental Institute, Universiti Sains Malaysia, Bertam, Penang, Malaysia

Correspondence Address:
Wan Adnan Wan Omar
Advanced Medical and Dental Institute, Universiti Sains Malaysia, Bertam, 13200 Kepala Batas, Pulau Pinang
Malaysia
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/pm.pm_226_19

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Aim/Background: In colorectal cancer, TAC1 was shown to be highly methylated in early carcinogenesis. Using HT29 cell line as a model of colorectal cancer, we postulated that Phyllanthus debilis methanolic extract could regulate DNA methylation in TAC1 gene promoter region and thus could alter the progression of colorectal cancer. Methodology: Cell culture study was done using HT29 cells, which were treated with 0.117 mg/ml P. debilis methanolic extract or 0.5 μM 5-Aza-2-Deoxycytidine (5-Aza) for 72 h. Cells were harvested at 72 h and were extracted for DNA. The DNA was bisulfite modified before been PCR and later was pyrosequenced. Results: The treatment with P. debilis significantly decreased the DNA methylation of TAC1 gene at site 1 (93.1% ± 2.4% vs. 100% ± 0.1%, P < 0.05), site 2 (86.5% ± 1.3% vs. 91.7% ± 0.2%, P < 0.05), and site 3 (96.7% ± 0.9% vs. 100% ± 0%, P < 0.05); but, no significant changes of DNA methylation were seen at site 4 (96.5% ± 1.9% vs. 93.6% ±0.6%, P > 0.05). The average of all Cytosine nucleotide followed by Guanine nucleotide (CpG) sites methylation was reduced, but not statistically significant when compared to the untreated cells (mean methylation 93.2% ± 2.4% vs. 96.3% ± 2.2%, P > 0.05). For cells treated with 5-Aza, DNA methylation was significantly decreased only at site 2 (88.6% ±0.8% vs. 91.7% ±0.2%, P < 0.05), but there was no significant methylation changes at site 1 (94.8% ± 2.4% vs. 100.0% ±0.06%, P > 0.05), site 3 (98.9% ± 1.1% vs. 100% ± 0%, P > 0.05), site 4 (90.6% ±1.6% vs. 93.6% ±0.6%, P > 0.05), and the average of all CpG sites (mean methylation 93.2% ± 2.3% vs. 96.3% ± 2.2%, P > 0.05). Conclusion: Treatment of methanolic extract of P. debilis reduces the methylation of promoter region of TAC1 gene, with better effect than low dose 5-Aza at 72 h of treatment. The anticancer effect of P. debilis may be partly been regulated through DNA methylation.


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