miR-324-5p inhibits gallbladder carcinoma cell metastatic behaviours by downregulation of transforming growth factor beta 2 expression
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Published on: October 27, 2020
Xinrong Zhang1, Lei Zhang2, Ming Chen3
1Department of Digestive Medicine, Tianjin Nankai Hospital, Tianjin, People's Republic of China.
Abstract:
Increasing studies have demonstrated that microRNAs (miRNAs) are associated with the metastasis of gallbladder carcinoma (GBC). Recently, miR-324-5p has been reported to be a tumour-suppressive miRNA in many types of malignant cancer. However, the biological function and molecular mechanism of miR-324-5p in GBC still remain largely unknown. Here, we found that miR-324-5p expression was notably down-regulated in both GBC tissues and cells compared with that in normal controls. Downregulated miR-324-5p expression was negatively associated with the status of local invasion and lymph node metastasis and predicted a poor prognosis in GBC patients. Further functional assays revealed that restoration of miR-324-5p significantly suppressed GBC cell migration, invasion and epithelial-mesenchymal transition (EMT) in vitro and impeded the metastasis of GBC cells in vivo. Moreover, RNA immunoprecipitation (RIP) and dual-luciferase reporter assay confirmed that the transforming growth factor beta 2 (TGFB2) was a direct target gene of miR-324-5p in GBC cells. Mechanically, small interfering RNA (siRNA)-mediated knockdown of TGFB2 partially phenocopied the inhibitory effects of miR-324-5p overexpression on GBC cell metastatic phenotypes. In summary, our findings demonstrated that miR-324-5p targets TGFB2 expression to inhibit GBC cell metastatic behaviors, and implying miR-324-5p as a potential biomarker for diagnostic and therapeutic strategies in GBC.
Insights
MicroRNA-324-5p (miR-324-5p) suppresses gallbladder carcinoma (GBC) metastasis by targeting TGFB2. Lower miR-324-5p levels indicate poor prognosis and are linked to GBC invasion and metastasis.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- MicroRNAs (miRNAs) play a role in gallbladder carcinoma (GBC) metastasis.
- miR-324-5p is a known tumor suppressor in various cancers, but its function in GBC is unclear.
Purpose of the Study:
- To investigate the role and mechanism of miR-324-5p in gallbladder carcinoma (GBC) metastasis.
Main Methods:
- Compared miR-324-5p expression in GBC and normal tissues/cells.
- Performed in vitro and in vivo functional assays to assess miR-324-5p's effect on GBC cell migration and invasion.
- Utilized RNA immunoprecipitation (RIP) and dual-luciferase reporter assays to identify miR-324-5p targets.
- Used siRNA to knockdown TGFB2 to validate its role.
Main Results:
- miR-324-5p was significantly downregulated in GBC tissues and cells.
- Downregulated miR-324-5p correlated with local invasion, lymph node metastasis, and poor patient prognosis.
- Restoring miR-324-5p inhibited GBC cell migration, invasion, epithelial-mesenchymal transition (EMT), and metastasis in vitro and in vivo.
- Transforming growth factor beta 2 (TGFB2) was identified as a direct target of miR-324-5p.
- TGFB2 knockdown partially mimicked the anti-metastatic effects of miR-324-5p.
Conclusions:
- miR-324-5p inhibits GBC cell metastasis by targeting TGFB2.
- miR-324-5p shows potential as a diagnostic biomarker and therapeutic target for GBC.

