Antioxidant and chemotherapeutic synergy: Triphala enhances doxorubicin cytotoxicity in breast cancer cells and reduces toxicity in non-tumorigenic cells

Supamas  Charucharana; Poonlarp  Cheepsunthorn; Chalisa  Louicharoen Cheepsunthorn

Authors

Supamas Charucharana Medical Sciences Program, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand and Faculty of Sciences and Technology, Phranakhon Rajabhat University, Bangkok, Thailand
Poonlarp Cheepsunthorn Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand
Chalisa Louicharoen Cheepsunthorn Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand

Keywords:

antioxidant therapy, breast cancer, chemotherapy, doxorubicin, gallic acid, reactive oxygen species, synergistic effect, triphala

Abstract

Background: The combination of antioxidants with chemotherapy is increasingly being explored to minimize toxicity in noncancerous cells during breast cancer treatment. Triphala (TPL), a Thai herbal compound rich in antioxidants, shows potential as a complementary candidate for use in breast cancer chemotherapy.

Objective: This study investigated the combined effects of TPL and low-dose doxorubicin (DOX) on human breast cancer and non-tumorigenic mammary epithelial cells.

Methods: TPL’s bioactive compounds were analyzed using high-performance liquid chromatography. Cell viability and the levels of reactive oxygen species (ROS) were assessed in breast cancer (MDA-MB-231, MCF-7) and epithelial (MCF-10A) cells using MTT and chloromethyl 22 ,72- dichlorodihydrofluorescein diacetate assays performed in triplicate. The combination index (CI) values were determined by CompuSyn. Furthermore, the mRNA expression of apoptosis- and antioxidant-related genes was evaluated using qPCR.

Results: Gallic acid (11.9%) was identified as the major component in TPL. The combination of TPL and lowdose DOX synergistically enhanced the cytotoxicity in MCF-7 and MDA-MB-231 cells. This combination significantly reduced the expression of the antioxidant genes SOD1 and GPX1 in MDA-MB-231 (SOD1: P < 0.001, GPX1: P = 0.035) and MCF-7 (SOD1: P = 0.035, GPX1: P = 0.036) cells, which resulted in increased ROS levels in MDA-MB-231 (P = 0.005) and MCF-7 (P = 0.008) cells. Elevated ROS triggered apoptosis via the increased BAX/BCL2 ratio in MDA-MB-231 (P < 0.001) and MCF-7 (P = 0.02) cells. Conversely, TPL displayed protective effects in nontumorigenic MCF-10A cells by upregulating SOD1 (P = 0.031) and GPX1 (P < 0.001), reducing ROS (P = 0.002), and lowering the BAX/BCL2 ratio (P < 0.001), thereby promoting cell survival.

Conclusion: TPL, in combination with low-dose DOX, effectively induces cytotoxicity in breast cancer cells while protecting non-tumorigenic cells, which suggests its potential as complementary therapy in breast cancer treatment.

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