Effect of Mono-2-ethyhexyl Phthalate on DNA Methylation in Human Prostate Cancer LNCaP Cells

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Objective To evaluate whether mono(2‐ethylhexyl) phthalate(MEHP) affects genomic DNA methylation and the methylation status of some specific genes such as patched gene(PTCH) and smoothened gene(SMO) in LNCaP cells. Methods LNCaP cells were treated with MEHP(0, 1, 5, 10, and 25 μmol/L) for 3 days. An ELISA assay was preformed to detect genomic methylation, including 5‐methylcytosine(5‐mC) and 5‐hydroxymethylcytosine(5‐hmC) content. A pyrosequencing assay was applied to assess DNA methylation in PTCH and SMO gene promoters. The correlation between DNA methylation and gene expression was assessed. Results The proportion of cytosines with 5‐mC methylation in LNCaP cells was significantly decreased by MEHP(1, 5, 10, and 25 μmol/L) in a dose‐dependent manner(P < 0.01). For genes in the Hedgehog pathway, there was no significant MEHP concentration‐dependent difference in the DNA methylation of PTCH and SMO. Conclusion MEHP might affect the progression of prostate cancer through its effect on global DNA methylation. Objective To evaluate mono (2-ethylhexyl) phthalate (MEHP) affects genomic DNA methylation and the methylation status of some specific genes such as patched gene (PTCH) and smoothened gene (SMO) in LNCaP cells. Methods LNCaP cells were treated with MEHP (0, 1, 5, 10, and 25 μmol / L) for 3 days. An ELISA assay was preformed to detect genomic methylation, including 5- methylcytosine (5-mC) and 5-hydroxymethylcytosine The correlation between DNA methylation and gene expression was assessed. Results The proportion of cytosines with 5-mC methylation in LNCaP cells was significantly decreased by MEHP (1, 5, 10 , and 25 μmol / L) in a dose-dependent manner (P <0.01). For genes in the Hedgehog pathway, there was no significant MEHP concentration-dependent difference in the DNA methylation of PTCH and SMO. Conclusion MEHP might affect the progression of prostate cancer thro ugh its effect on global DNA methylation.
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