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Fig. 2 | Clinical Epigenetics

Fig. 2

From: A multiplatform approach identifies miR-152-3p as a common epigenetically regulated onco-suppressor in prostate cancer targeting TMEM97

Fig. 2

COPZ2-miR-152-3p transcriptional unit’s DNA methylation and expression validation in IPO Porto’s cohort of prostate samples. a HumanMethylation450 BeadChip data for miR-152-3p locus, showing a significant increase in cg21384971, cg05096161, cg05850656, cg06598332, cg10382221, and cg24389730. b Significant miR-152-3p downregulation in PCa (n = 100) compared to morphologically normal prostate tissues (MNPT, n = 14), as determined by RT-qPCR (p < 0.0001). c Significantly decreased COPZ2 transcript levels in PCa samples (p = 0.0022). d COPZ2-miR-152-3p promoter hypermethylation in PCa samples determined by pyrosequencing (p = < 0.0001). e MiR-152-3p expression levels in PCa cell lines compared to benign RWPE cells [expression (E):1] assessed by RT-qPCR. Expression is significantly lower in LNCaP (E ~ 0.13), DU145 (E ~ 0.54) and PC3 (E ~ 0.09) cells. f Significantly lower COPZ2 expression levels by RT-qPCR in LNCaP (E ~ 0.005), DU145 (E ~ 0.17) and PC3 (E ~ 0.12) cells compared to RWPE (E 1). g Prostate cell lines’ DNA methylation profiling. LNCaP (FC of methylation levels: ~ 59) and PC3 (FC: ~ 7) cells showed increased miR152-COPZ2 promoter hypermethylation compared to RWPE (FC: 1), 22RV1 and DU145 cells. h A 72-h 5-Aza-CdR exposure, associated with significant decrease in promoter methylation levels of the transcriptional unit COPZ2-miR-152-3p in LNCaP (decreased ~ 40%) and PC3 (decreased ~ 68%) cells. LNCaP and PC3 cells miR-152-3p (i) and COPZ2’s (j) expression levels following 72-h exposure to 5-Aza-CdR associated with increased miR-152-3p expression levels (FC:1.75; 1.95, respectively) and COPZ2 (FC:1.12; 3.98, respectively). Error bars represent the s.d. for three biological replicates. Mann-Whitney U test: *p < 0.05, **p < 0.01, ***p < 0.001. β values range from 0 to 1 and were determined by the ratio of the methylated probe intensity and the overall intensity (sum of methylated and unmethylated probe intensities). MNPT: Morphologically Normal Prostate Tissue; PCa: Prostate Cancer; 5-Aza-CdR: 5-aza-2-deoxycytidine

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