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)

Abstract

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