Journal: Molecular Cancer Research
Article Title: Gut Microbiota–Mediated hsa_circ_0126925 Targets BCAA Metabolic Enzyme BCAT2 to Exacerbate Colorectal Cancer Progression
doi: 10.1158/1541-7786.MCR-24-0434
Figure Lengend Snippet: HFD-modulated gut microbiota regulates the expression of circRNAs. A, The circRNA expression profiles of intestinal tumors from CTL, HFD, and HFD+Abx AOM/DSS model mice were compared via high-throughput circRNA microarray sequencing. B, Mmu_circ_017841, mmu_circ_006689, mmu_circ_32011, circ_38945, mmu_circ_19191, mmu_circ_41893, and mmu_circ_45525, which appeared simultaneously in the upregulated circRNAs of the HFD vs. CTL and HFD+Abx groups. C–E , RT-qPCR was used to verify the differential expression of target mmu_circRNAs (mmu_circ_38945, mmu_circ_32011, mmu_circ_19191, and mmu_circ_45525) in groups of mice generated via three different modeling methods. F, Sanger sequencing results for circ_38945 and circ_0126925. G, Divergent and convergent primers were used to amplify circ_0126925, gDNA, and cDNA with or without RNase treatment before RT-qPCR. H, FISH analysis of the subcellular localization of circ_0126925. I, The proportions of cells in which circ_0126925 was expressed in the nucleus and cytoplasm. J and K, RT-qPCR and ISH were used to detect the expression of circ_0126925 in cancer and adjacent tissues. L, KM curve of the overall survival rate of patients with different expression levels of circ_0126925. In C–E , J , and K , unpaired and paired t tests were used for statistical analyses. In K , the differences between the KM curves were tested for significance by the log-rank test. *, P < 0.05; **, P < 0.01; ***, P < 0.001. ns indicates no significant difference between the two indicated groups.
Article Snippet: Then, the labeled circRNA was hybridized onto an Arraystar Mouse circRNA Array (8 × 15 K, Arraystar) and incubated in an Agilent Hybridization Incubator (Agilent) at 65°C for 17 hours.
Techniques: Expressing, High Throughput Screening Assay, Microarray, Sequencing, Quantitative RT-PCR, Quantitative Proteomics, Generated
