Modifications of The Human Liver Cancer Cells through microRNA-145-Mediated Targeting of CDCA3

OBJECTIVE: MicroRNA-143 (miR-145) has shown promising tumor-suppressive effects in different types of cancer and could potentially act as a tumor suppressor in liver cancer. To explore this further, the present study aims to investigate the protective function of miR-145 in human liver cancer. MATERIALS AND METHODS: In this experimental study, we analyzed miR-145 expression in human tissue samples and liver cancer cell lines using quantitative real time polymerase chain reaction (qRT-PCR). Cancer cell lines were transfected using Lipofectamine 2000. We assessed cell viability of HepG2 liver cancer cell line using the MTT assay and analyzed colony forming potential through clonogenic assay. Flow cytometry was employed to evaluate cell cycle phase distribution in cancer cell lines expressing either miR-145 inhibitor or miR-145 mimics. The motility of cancer cell lines was determined using the transwell chamber assay. Protein expression levels of Cyclin B1 and CDCA3, important for cell cycle progression and cell division, respectively, were measured using western blotting. Finally, we conducted a dual luciferase assay to investigate the interaction between miR-145 and CDCA3. RESULTS: Downregulation of miR-145 was observed in liver cancer cells. Overexpression of miR-145 through transfection inhibited cancer cell proliferation, while transfection of miR-145 inhibitor increased proliferation rate. MiR- 145 overexpression led to cell cycle arrest at the G2/M phase by suppressing cyclin B1 protein expression. Moreover, miR-145 overexpression suppressed migration and invasion of cancer cells. CDCA3 gene was identified as the intracellular target of miR-145, and the inhibitory effects of miR-145 were mediated through the CDCA3 protein. CONCLUSION: MiR-145 regulates CDCA3 in liver cancer, affecting its development. Decreased miR-145 allows CDCA3 accumulation, promoting liver cancer progression. MiR-145 targets CDCA3 to inhibit viability, migration, and invasion of liver cancer cells. Further research is needed to understand miR-145's regulatory role and develop more effective strategies against liver cancer.