Exosome-Derived Circ_0094343 Promotes Chemosensitivity of Colorectal Cancer Cells by Regulating Glycolysis via the miR-766-5p/TRIM67 Axis.

Abstract

Objective Currently, the role of circ_0094343 (circPTEN) on the chemosensitivity of CRC remains to be clarified. This study aimed to investigate the role and mechanism of exosome-delivered circ_0094343 in the proliferation, glycolysis, and chemosensitivity of colorectal cancer (CRC) cells. Methods Real-time quantitative polymerase chain reaction (qRT-PCR) was utilized to detect the expression level of circ_0094343, miR-766-5p, and TRIM67 (Tripartite motif-containing 67) in CRC clinical tissue samples and cells, transmission electron microscopy (TEM) to observe the morphology of exosomes, and nanoparticle tracking analysis (NTA) system to measure the diameter of exosomes. Besides, PKH67 fluorescent labeling was applied for assessing the level of exosome uptake by cells, MTT and cell clone formation assays for detecting cell proliferation and clone formation, respectively, and related kits for checking the glucose consumption, lactate production, and extracellular acidification rate (ECAR) in cells. Dual-luciferase reporter (DLR) gene assay was used for verifying the targeting relationship between circ_0094343 and miR-766-5p, miR-766-5p and TRIM67, RNA immunoprecipitation (RIP) experiment for the interaction between circ_0094343 and miR-766-5p, and Western blot for the protein level of exosome surface antigens (HSP70, CD63) and TRIM67 in cells in exosomes and cell lysates. Results circ_0094343 was significantly downregulated in CRC tissues, chemotherapy-resistant CRC tissues, and metastatic CRC tissues. Moreover, exosomes-carried circ_0094343 played an inhibitory role in the proliferation, clone formation and glycolysis of HCT116 cells. Meanwhile, it could also improve the chemosensitivity of HCT116 cells to 5-fluorouracil (5-FU), oxaliplatin (L-OHP), and doxorubicin (Dox). Additionally, circ_0094343 acted as a sponge for miR-766-5p, and miR-766-5p targeted and regulated TRIM67. In CRC tissues, miR-766-5p expression was negatively correlated with TRIM67 expression, while circ_0094343 was positively associated with TRIM67. Further, mechanistic validation also demonstrated that circ_0094343 could inhibit HCT116 cell proliferation, clone formation, glycolysis, and chemotherapy resistance via the miR-766-5p/TRIM67 axis. Conclusion circ_0094343 inhibited the proliferation, clone formation and glycolysis of CRC cells and improved their chemosensitivity to various chemotherapeutic drugs via the miR-766-5p/TRIM67 axis. This finding may provide new insights into the treatment of CRC.