Role of hsa_Circ_0001821 in Colorectal Cancer Pathogenesis and Response to 5-Fluorouracil through miR-203a-3p/FGF-2 Axis

Abstract

Background: Chemoresistance, the primary cause of disease relapse and treatment failure, poses a significant challenge in the treatment of colorectal cancer (CRC). Understanding the molecular mechanisms that underlie the pathogenesis and chemoresistance of colorectal tumor cells, as well as identifying novel therapeutic strategies, would be crucial. This study aimed to evaluate the role of hsa_Circ_0001821 in response to 5-fluorouracil (5-FU) in CRC, a topic that has not been examined to date.

Methods: The current study investigated the effect of hsa_Circ_0001821 suppression using interfering RNAs on the response of colorectal tumor cells to 5-FU. The expression levels of hsa_Circ_0001821, hsa-miR-203a-3p, BAX, BCL-2, and FGF-2 were determined via quantitative RT-PCR. Cell survival, migration rate, and apoptosis induction of colorectal tumor cells subjected to 5-FU treatment were assessed using the MTT test, scratch assay, and flow cytometry analysis, respectively.

Results: Knockdown of hsa_Circ_0001821 with siRNA increased the expression level of hsa-miR-203a-3p and decreased the expression level of FGF-2. Additionally, the knockdown of hsa_Circ_0001821 enhanced the sensitivity of colorectal tumor cells to 5-FU. This circRNA significantly affected the viability, apoptosis, and migration of tumor cells.

Conclusion: Our study reveals the potential role of hsa_Circ_0001821 in controlling the tumor cell viability and response to 5-FU by targeting the hsa-miR-203a-3p/FGF-2 axis. These findings enhance our understanding of the molecular mechanisms that influence chemotherapy response in CRC, paving the way for the identification of more effective treatments for this disease.

Keywords: Antineoplastic Agents; Circular RNA; Colorectal Neoplasm; MicroRNAs.

Fig 1

The expression level of (A) hsa-miR-203a-3p, (B) hsa_Circ_0001821, and (C) FGF-2 following the hsa_Circ_0001821 knockdown in HCT116 cell line. The data show that by suppressing hsa_Circ_0001821, hsa-miR-203a-3p is upregulated, while the expression level of FGF-2 genes is significantly downregulated (^*p < 0.05, ^**p < 0.01). ns: not significant

Fig 2

Effects of hsa_Circ_0001821 knockdown on CRC cell viability. The effect of hsa_Circ_0001821 knockdown was evaluated by MTT assay. The results indicated that the use of Si-Circ-0001821 led to a significant decrease in cell viability in HCT116 cells.

Fig. 3

Effect of hsa_Circ_0001821 silencing on the migration of HCT116 cells. (A) Representative optical microscopy images of the scratch assay used to determine the rate of cell migration. (B) The data show that in the group treated with Si-Circ-0001821 and 5-FU, the amount of cell migration was significantly inhibited compared to the control group (^*p < 0.05). ns: not significant

Fig 4

The hsa_Circ_0001821 silencing induced apoptosis in CRC cells. (A) The flow cytometry results indicated that the silencing hsa_Circ_0001821 could significantly induce apoptosis. Relative expression of (B) BAX and (C) BCL-2 genes was evaluated with real-time PCR and indicated that treatment of HCT116 cells with Si-Circ-0001821 and 5-Fu resulted in increasing the expression level of BAX and decreasing the expression level of BCL-2 (^*p < 0.05, ^**p < 0.01, ^***p < 0.001, and ^****p < 0.0001). (D) Dot blot diagrams were obtained by flow cytometry method. ns: not significant