Abstract
Signal transducer and activator of transcription 3 (STAT3), a transcriptional factor involved in tumorigenesis and cancer stemness formation, contributes to drug resistance in cancer therapies. STAT3 not only mediates gene transcription but also participates in microRNA suppression. This study identified a STAT3-downstream micro RNA (miRNA) involved in drug resistance against regorafenib in colorectal cancer stem-like tumorspheres. Small RNAseq was used to investigate differential microRNAs in colorectal cancer cell-derived tumorspheres and in a STAT3-knockdown strain. The miRNA-mediated genes were identified by comparing RNAseq data with gene targets predicted using TargetScan. Assays for detecting cell viability and apoptosis were used to validate findings. The formation of colorectal cancer stem-like tumorspheres was inhibited by BBI608, a STAT3 inhibitor, but not by regorafenib. Additional investigations for microRNA expression demonstrated an increase in 10 miRNAs and a decrease in 13 miRNAs in HT29-derived tumorspheres. A comparison of small RNAseq results between tumorspheres and HT29shSTAT3 cells revealed the presence of four STAT3-mediated miRNAs in HT29-derived tumorspheres: hsa-miR-215-5p, hsa-miR-4521, and hsa-miR-215-3p were upregulated, whereas miR-30a-5p was downregulated. Furthermore, hsa-miR-4521 was associated with poor overall survival probability, and miR-30a-5p was associated with better overall survival probability in patients with rectum cancer. Comparisons of RNAseq findings between HCT116- and HT29-derived tumorspheres revealed that HSPA5 were mediated by the STAT3-miR-30a-5p axis, which is overexpressed in colorectal tumorspheres associating to anti-apoptosis. In addition, the transfection of miR-30a-5p and inhibition of HSPA5 by HA15 significantly reduced cell viability and increased apoptosis in HT29 cells. In conclusion, a STAT3-miR-30a-5p-HSPA5 axis was observed against regorafenib-mediated apoptosis in colorectal cancer tumorspheres. The expression of miR-30a-5p was repressed by STAT3; in addition, HSPA5 was identified as the target gene of miR-30a-5p and contributed to both tumorsphere formation and anti-apoptosis.
Highlights
Colorectal carcinoma (CRC) is a lethal disease with a high prevalence worldwide; it is reported as the third most common cancer globally with a death rate of approximately 10% [1]
We investigated the role of signal transducer and activator of transcription 3 (STAT3) in drug resistance against regorafenib and determined the mechanism involved
HCC116-and HT29-derived tumorspheres were cultured in serum-free Dulbecco’s modified Eagle’s medium (DMEM) medium containing individual 20 ng/mL of epidermal growth factor (EGF), basic fibroblast growth factor, hepatocyte growth factor (HGF), and interleukin (IL)-6, which are sensitive to STAT3 inhibitors [9]
Summary
Colorectal carcinoma (CRC) is a lethal disease with a high prevalence worldwide; it is reported as the third most common cancer globally with a death rate of approximately 10% [1]. Regorafenib could increase signal transducer and activator of transcription 3 (STAT3) phosphorylation in colorectal cancer cells, HCT116 and HT29 [3]. Another study reports that regorafenib significantly inhibited intrinsic STAT3 phosphorylation in hepatocellular carcinoma cell lines due to increased SHP-1 phosphatase activity, resulting in cell apoptosis [4]. Regorafenib inhibits the activation of receptor tyrosine kinases (RTKs), including FGFR, PDGFR, VEGFR, TIE-2, RET, and KIT, resulting in a reduction in downstream signaling [7]. Based on these findings, we assumed that STAT3, induced by regorafenib under stressful conditions in CD133-positive HCT116 and HT29 cells, plays a vital role in maintaining both tumor survival and cancer stemness. The miRNA expression profiles between HT29-derived stem-like tumorspheres and HT29shSTAT3 cells were compared
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
