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MicroRNA-520c-3p Modulates Doxorubicin-Chemosensitivity in HepG2 Cells
Mohamed A Ragheb1, Marwa H Soliman1, Emad M Elzayat2
1Department of Chemistry (Biochemistry Division), Faculty of Science, Cairo University, Giza, Egypt.
Background:
Doxorubicin (DOX) is one of the most common drugs used in cancer therapy, including Hepatocellular Carcinoma (HCC). Drug resistance is one of chemotherapy's significant problems. Emerging studies have shown that microRNAs (miRNAs) could participate in regulating this mechanism. Nevertheless, the impact of miRNAs on HCC chemoresistance is still enigmatic.
Objective:
Investigating the role of microRNA-520c-3p (miR-520c-3p) in the enhancement of the anti-tumor effect of DOX against HepG2 cells.
Methods:
Expression profile for liver-related miRNAs (384 miRNAs) has been analyzed on HepG2 cells treated with DOX using qRT-PCR. miR-520c-3p, the most deregulated miRNA, was selected for combination treatment with DOX. The expression level for LEF1, CDK2, CDH1, VIM, Mcl-1 and p53 was evaluated in miR-520c-3p transfected cells. Cell viability, colony formation, wound healing as well as apoptosis assays have been demonstrated. Furthermore, Mcl-1 protein level was measured using the western blot technique.
Results:
The present data indicated that miR-520c-3p overexpression could render HepG2 cells chemo-sensitive to DOX through enhancing its suppressive effects on proliferation, migration, and induction of apoptosis. The suppressive effect of miR-520c-3p involved altering the expression levels of some key regulators of cell cycle, proliferation, migration and apoptosis, including LEF1, CDK2, CDH1, VIM, Mcl-1 and p53. Interestingly, Mcl-1 was found to be one of the potential targets of miR-520c-3p, and its protein expression level was down-regulated upon miR-520c-3p overexpression.
Conclusion:
Our data referred to the tumor suppressor function of miR-520c-3p that could modulate the chemosensitivity of HepG2 cells towards DOX treatment, providing a promising therapeutic strategy in HCC.
Insights
MicroRNA-520c-3p enhances doxorubicin's anti-cancer effects in liver cancer cells by increasing sensitivity and promoting apoptosis. This microRNA (miRNA) targets Mcl-1, offering a potential new strategy for hepatocellular carcinoma (HCC) treatment.
Area of Science:
- Oncology
- Molecular Biology
- Biochemistry
Background:
- Doxorubicin (DOX) is a key chemotherapy drug for Hepatocellular Carcinoma (HCC).
- Drug resistance significantly hinders chemotherapy efficacy.
- MicroRNAs (miRNAs) are implicated in chemoresistance, but their precise role in HCC remains unclear.
Purpose of the Study:
- To investigate the role of microRNA-520c-3p (miR-520c-3p) in modulating DOX chemoresistance in HepG2 HCC cells.
- To assess the impact of miR-520c-3p on DOX's anti-tumor activity.
Main Methods:
- Quantitative real-time PCR (qRT-PCR) to profile miRNA expression in DOX-treated HepG2 cells.
- Transfection of HepG2 cells with miR-520c-3p for combination therapy studies.
- Assays for cell viability, colony formation, migration, and apoptosis.
- Western blotting to measure Mcl-1 protein levels.
Main Results:
- Overexpression of miR-520c-3p enhanced DOX's anti-tumor effects, increasing chemo-sensitivity in HepG2 cells.
- miR-520c-3p modulated key regulators of cell cycle, proliferation, migration, and apoptosis (LEF1, CDK2, CDH1, VIM, Mcl-1, p53).
- Mcl-1 was identified as a target of miR-520c-3p, with its protein expression reduced upon miR-520c-3p overexpression.
Conclusions:
- miR-520c-3p exhibits tumor suppressor functions by enhancing HepG2 cell chemosensitivity to DOX.
- This microRNA holds promise as a therapeutic strategy to overcome DOX resistance in HCC.
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