Synergistic Action of 1,2-Epoxy-3 (3- (3,4-dimethoxyphenyl)-4H-1-benzopiyran-4-on) Propane with Doxorubicin and Cisplatin through Increasing of p53, TIMP-3, and MicroRNA-34a in Cervical Cancer Cell Line (HeLa)

Document Type: Research Articles

Authors

1 Department of Biochemistry, Faculty of Medicine Universitas Islam Bandung, Bandung, Indonesia.

2 Doctorate Program, Faculty of Medicine, Public Health and Nursing, Universitas Gadjah Mada, Yogyakarta, Indonesia.

3 Department of Pharmacology and Therapy, Faculty of Medicine, Public Health and Nursing, Universitas Gadjah Mada, Yogyakarta, Indonesia.

4 Department of Surgery, Faculty of Medicine, Public Health and Nursing, Universitas Gadjah Mada, Yogyakarta, Indonesia.

5 Department of Histology, Faculty of Medicine, Public Health and Nursing, Universitas Gadjah Mada, Yogyakarta, Indonesia.

Abstract

Objective: Cervical cancer is the second most common cancer among women worldwide, with a high mortality rate
especially in developing countries. Insufficient treatment for cervical cancer, multiple side effects, and high drug prices
encourage researchers to look for effective and selective cancer drugs with appropriate molecular targets. This study
explored the cytotoxicity of (1,2-epoxy-3(3-(3,4-dimethoxyphenyl)-4H-1-benzopyran-4-on) propane (EPI) synthesized
from clove leaves oil on HeLa cells, its combination with doxorubicin (DOX) and cisplatin (CIS), and also their influence
on p53, TIMP-3, and miR-34a as therapeutic targets. Materials and Methods: This research was an experimental
in vitro study on cervical cancer uteri culture. The cytotoxicity was analyzed by MTT assay. The drug combination
synergisms were indicated by the combination index (CI) (using CompuSyn 1.4). HeLa cells in 32 wells were divided
into eight groups as negative control, which were given EPI ½IC50, EPI IC50, EPI 2IC50, DOX IC50, combination of
EPI+DOX, CIS, and the combination of EPI+CIS. The p53 and TIMP-3 concentrations were measured using ELISA,
and expressions of miR-34a with qRT-PCR. One-way ANOVA and post hoc Tukey tests were performed to determine
the mean difference of all variables between the study groups. Results: IC50 for EPI was 33.24 (±3.01) μg/ml, while DOX
and CIS were 4.8 μg/ml (±0.1), and 23.34 μg/ml (±3.01), respectively, while CI values for EPI-DOX were EPI-CIS and 8 (2.93±0.42) μg/ml were significantly higher compared to the control group (p<0.05). All treatment groups showed
significantly increased miR-34a expressions compared to the control group (p<0.05). Conclusion: The combinations
showed a very strong synergism and a moderate slight synergism for EPI-DOX and EPI-CIS. Both combinations were
able to increase the expressions of p53, TIMP-3 proteins, and MiR-34a in the HeLa cells.

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