Research Paper Volume 12, Issue 21 pp 21404—21422

Figure 4. CPEB3 directly targets the 3’UTR of JAK1 and regulates colorectal cancer proliferation and metastasis via CPEB3/JAK/STAT axis. (A) High-throughput sequencing analysis of mRNAs bound to CPEB3 in SW480 cells showed the top 10 significantly enriched terms using Gene ontology analysis. (B) RNA immunoprecipitation and high-throughput sequencing analysis showed that JAK1 mRNA was significantly enriched in CPEB3 immunoprecipitates. (C) The combination of JAK1 mRNA and CPEB3 was further validated by RIP-qPCR. IgG and SnRNP70 are used as the negative and positive control respectively. Data are shown as mean ± SD (***P< 0.001). (D) Luciferase reporter plasmid appended with different 3’UTR fragments of JAK1 mRNA were co-transfected with CPEB3 in the HEK-293T cell line, and luciferase activity was detected. Data are displayed as the means ± SD (**P< 0.01, NS: No Significance). (E, F) Impairment of JAK/STAT pathway by Ruxolitinib could reverse the effect of CPEB3 down-regulation on proliferation ability by CCK-8 assay (E) as well as the migration and invasion ability by transwell assays (original magnification, ×400) of colorectal cancer cell lines. (F).