Cytotoxic and Antimetastatic Activity of Hesperetin and Doxorubicin Combination Toward Her2 Expressing Breast Cancer Cells

Ika Putri Nurhayati,Annisa Khumaira,Edy Meiyanto,Adam Hermawan,Gagas Pradani Nur Ilmawati

doi:10.31557/apjcp.2020.21.5.1259

Abstract

Objective:This study aimed to explore Hesperetin (Hst) potency as a co-chemotherapeutics agent combined with Doxorubicin (Dox), particularly cytotoxic and antimetastasis effects toward MCF-7/HER2 cells. Methods:The cytotoxic effects were measured under MTT assay. The flowcytometry analysis was used to examine the cell cycle modulation and apoptosis evidence, while the effect of migration was assayed by scratch wound healing assay. Western blotting and gelatin zymography were carried out to examine the expression level of proteins, HER2, and Rac1. Results:Under MTT assay, Hst and Dox exhibited to decrease cell viability in a dose-dependent manner with the IC50 value of 377 and 0,8 µM, respectively. The combination of Hst and Dox at the respective doses of 95 and 0,2 µM showed a synergistic effect with the combination index of 0,63. Flow cytometry analysis of Hst-Dox revealed that those compounds caused cell cycle arrest at the G2/M phase and induced apoptosis. Hst also decreased HER2 and Rac1 expression, as shown by western blot. Hst inhibited lamellipodia formation and cell migration, as indicated by microscopic observation and wound healing scratch assay. The antimetastatic activity of Hst was associated with the reduction of Rac1 and MMP9 expression as measured by gelatine zymography assay. Conclusion:These results indicated that the combination of Hst and Dox-induced cell cycle arrest, apoptosis, decreased HER2, Rac1, MMP9 expression, and cell migration. Thus, Hst may have the potential to be developed as a co-chemotherapeutic agent combined with doxorubicin toward HER2 overexpressing breast cancer cells.

Highlights

HER2 is one of epidermal growth factor receptor family that initiates various signaling pathways, including cell proliferation, survival, differentiation, angiogenesis, and invasion
This study aimed to explore Hesperetin (Hst) potency as a co-chemotherapeutics agent combined with Doxorubicin (Dox), cytotoxic and antimetastasis effects toward MCF-7/HER2 cells
These results indicated that the combination of Hst and Dox-induced cell cycle arrest, apoptosis, decreased HER2, Rac1, MMP9 expression, and cell migration

Summary

Introduction

HER2 is one of epidermal growth factor receptor family that initiates various signaling pathways, including cell proliferation, survival, differentiation, angiogenesis, and invasion. HER2 amplification leads to HER2 overexpression, which found in various types of cancers, such as breast, gastric, esophageal, ovarian, and endometrial cancers (Iqbal and Iqbal, 2014). HER2 overexpression promotes chemoresistance and increases metastasis potency (De Potter, 1994; Knuefermann et al, 2003). HER2 becomes one of the targeted therapy in cancer. Some efforts in this issue have been attempted and resulted in some new findings, such as antibody and small molecules targeted-HER2, but it remains some disadvantages, such as developing resistance of cancer and relatively expensive. The new chemotherapeutic agents targeted to HER2 still need to be developed

Methods

Results

Conclusion