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Fig. 6 | Molecular Cancer

Fig. 6

From: The circSPON2/miR-331-3p axis regulates PRMT5, an epigenetic regulator of CAMK2N1 transcription and prostate cancer progression

Fig. 6

Expression profiles of circRNAs and clinical features of circSPON2 in PCa. A A cluster heap map presented the significantly dysregulated circRNAs in human PCa tissues relative to adjacent normal tissues. Red represents upregulation and green represents downregulation. B Scatter plot of differentially expressed circRNAs in PCa and adjacent normal tissues. The X- and Y-axes represent average signal values of circRNAs from PCa and adjacent normal tissues. C Volcano plot of differentially expressed circRNAs. The values of X- and Y-axes in the volcano plot are the fold change (log2 transformed) and P value between PCa and adjacent normal tissues, respectively. Red/Blue dots indicate twofold change differentially expressed genes with statistical significance (142 blue dots, downregulated circRNAs in PCa tissues, and 288 red dots, upregulated circRNAs in PCa tissues). Yellow dots indicate non-differentially expressed circRNAs in PCa tissues. D Schematic illustration of circSPON2 formation from SPON2 gene in chromosome 4. E Relative expression of circSPON2 was determined in human prostate epithelial cell line RWPE-1, and PCa cell lines (LNCaP, PC-3, 22RV1, DU145 and VCaP) by qRT-PCR. F The existence of circSPON2 was confirmed by RT-PCR and gel electrophoresis in PC-3 cells. The back splicing junction of circSPON2 was verified by Sanger sequencing. G qRT-PCR analysis for the circSPON2 and SPON2 mRNA using the template cDNA reverse-transcribed by random primers and oligo dT primers using RNAs from PC-3 and DU145 cells. H Stability of circSPON2 and SPON2 mRNA was assessed by RNase R treatment followed by qRT-PCR. I qRT-PCR assay for the expression of circSPON2 and SPON2 mRNA in PC-3 cells treated with Actinomycin D (2 μg/mL) at the indicated time points. J Fluorescence in situ hybridization (FISH) with Cy3-labeled circSPON2 probes (red) were performed to detect the location of circSPON2 in PC-3 cells. Scale bar = 20 μm. K Comparison of circSPON2 levels between PCa and paired adjacent normal tissues (n = 248). L Comparison of circSPON2 levels in different Gleason scores (GS) were analyzed in PCa tissues (n = 248). M Comparison of circSPON2 levels in different T stages were analyzed in PCa tissues (n = 248). N Kaplan–Meier curve was used to evaluate the progression-free survival (PFS) of PCa patients with high (n = 124) or low (n = 124) circSPON2 expression (Log-rank, P < 0.05). Data were showed as mean ± SD. *P < 0.05, **P < 0.01

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