Dietary isoflavone daidzein synergizes centchroman action via induction of apoptosis and inhibition of PI3K/Akt pathway in MCF-7/MDA MB-231 human breast cancer cells

Shweta Kaushik; Hari Shyam; Ramesh Sharma; Anil K Balapure

doi:10.1016/j.phymed.2018.01.007

Dietary isoflavone daidzein synergizes centchroman action via induction of apoptosis and inhibition of PI3K/Akt pathway in MCF-7/MDA MB-231 human breast cancer cells

Phytomedicine. 2018 Feb 1:40:116-124. doi: 10.1016/j.phymed.2018.01.007. Epub 2018 Jan 31.

Authors

Shweta Kaushik¹, Hari Shyam², Ramesh Sharma², Anil K Balapure³

Affiliations

¹ Division of Biochemistry, CSIR-Central Drug Research Institute, Lucknow 226031, India; Academy of Scientific and Innovative Research (AcSIR), Taramani, Chennai, Tamil Nadu 600113, India.
² Division of Biochemistry, CSIR-Central Drug Research Institute, Lucknow 226031, India.
³ Division of Biochemistry, CSIR-Central Drug Research Institute, Lucknow 226031, India; Academy of Scientific and Innovative Research (AcSIR), Taramani, Chennai, Tamil Nadu 600113, India. Electronic address: anilbalapure58@gmail.com.

PMID: 29496164
DOI: 10.1016/j.phymed.2018.01.007

Abstract

Background: Despite advancements in the prognosis and management of breast cancer, it remains a major cause of mortality in women worldwide. Centchroman (CC), an oral contraceptive has been found to exhibit anti-cancer potential against a wide range of cancer including breast cancer.

Purpose: The present study is intended to evaluate the ability of soy isoflavone Daidzein (DZ) in enhancing the efficacy of CC in Human Breast Cancer Cells (HBCCs).

Methods/study design: Sulforhodamine B assay was employed to determine the cytotoxicity induced by 10 µM CC & 50 µM DZ separately and together in MCF-7/MDA MB-231 HBCCs and non-tumorigenic Human Mammary Epithelial Cells (HMECs) MCF-10A as a control. Combination Index (CI) analysis was executed using CompuSyn software. Further, apoptosis was assessed using Annexin V/PI, AO/PI staining and tunel assay. Cell cycle, reactive oxygen species generation and mitochondrial membrane potential alteration was determined using flow cytometry. Western blot analysis was performed to check the expression of respective proteins.

Results: The results suggest that the combination exerts elevated toxicity as compared to control and each drug per se without affecting HMECs MCF-10A. This therefore implies cancer cell specific action of CC plus DZ administered together. Additionally, combination index analysis suggests synergistic action of CC and DZ combination in HBCCs. Cell cycle analysis, Annexin V/PI staining, tunel assay and western blot analysis confirms the induction of apoptosis by combination in HBCCs. Interestingly, western blot analysis also revealed that the combination down-regulated the expression of proteins involved in cell survival i.e. PI3K, Akt and mTOR, suggesting inhibition of cell survival pathway.

Conclusion: The results overall demonstrate that CC plus DZ has higher anticancer efficacy as compared to either drug alone. Hence, the combination of CC plus DZ may offer a novel strategy for the management of breast cancer.

Keywords: Apoptosis; Centchroman; Daidzein; Human breast cancer cells; PI3K/Akt.

MeSH terms

Antineoplastic Combined Chemotherapy Protocols / pharmacology*
Apoptosis / drug effects
Breast Neoplasms / drug therapy*
Breast Neoplasms / metabolism*
Breast Neoplasms / pathology
Cell Cycle / drug effects
Cell Line, Tumor
Cell Survival / drug effects
Centchroman / administration & dosage
Centchroman / pharmacology
Drug Synergism
Female
Humans
Isoflavones / administration & dosage
Isoflavones / pharmacology
MCF-7 Cells
Membrane Potential, Mitochondrial / drug effects
Phosphatidylinositol 3-Kinases / metabolism*
Proto-Oncogene Proteins c-akt / metabolism*
TOR Serine-Threonine Kinases / metabolism

Substances

Isoflavones
Centchroman
daidzein
MTOR protein, human
Proto-Oncogene Proteins c-akt
TOR Serine-Threonine Kinases