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Fig. 1 | Molecular Cancer

Fig. 1

From: A novel protein encoded by circINSIG1 reprograms cholesterol metabolism by promoting the ubiquitin-dependent degradation of INSIG1 in colorectal cancer

Fig. 1

Characterization and clinical signification of hypoxia-induced circINSIG1 in CRC. A Differentially expressed circRNAs under hypoxia by circRNAs sequencing in HCT8, HCT116 and DLD1 cells. B Heatmap of the expression of 18 circRNAs in HCT8, HCT116 and DLD1 cells under hypoxia. C qRT-PCR analysis of circINSIG1 expression in CoCl2-induced cellular pseudo-hypoxia models. D qRT-PCR analysis of circINSIG1 expression in 85 paired CRC samples and normal adjacent tissues. E Comparison of circINSIG1 expression between patients with T stage 3–4 (n = 53) and those with T stage 1–2 (n = 32), detected by qRT-PCR. F Comparison of circINSIG1 expression between patients with clinical stage III–IV (n = 35) and those with clinical stage I–II (n = 50), detected by qRT-PCR. G Genomic loci of the INSIG1 gene and circINSIG1. H Red arrow indicates the back-splicing of INSIG1 exon 3 and 4 confirmed by Sanger sequencing. I RT-PCR analysis of the existence of circINSIG1 with the divergent primers and convergent primers in complementary DNA (cDNA) and genomic DNA (gDNA). J qRT-PCR analysis of the abundance of circINSIG1 and INSIG1 linear mRNA in HCT8 cells treated with actinomycin D at the indicated time points. K qRT-PCR analysis of circINSIG1 and INSIG1 linear mRNA treated with or without RNase R. L qRT-PCR analysis of circINSIG1 location in the nucleus or cytoplasm in HCT8 and DLD1 cells. GADPH served as a marker of cytoplasmic location, while U6 served as a marker of nuclear location. M Representative FISH images of circINSIG1 staining in HCT8 cells. Scale bar = 10 μm. N Representative images of ISH circINSIG1 expression in the paraffin-embedded CRC tissues and paired normal adjacent tissues. Scale bar = 250 μm and 50 μm respectively. O Kaplan-Meier curves for survival of CRC patients with low vs. high expression of circINSIG1. N, normal adjacent tissues; T, tumor tissues. Values are represented as mean ± SD. **p < 0.01, ***p < 0.001; ns, no significance, by 2-tailed Student’s t test

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