Bioinformatics and In vitro Studies Reveal the Importance of p53, PPARG and Notch Signaling Pathway in Inhibition of Breast Cancer Stem Cells by Hesperetin

Abstract

Purpose: The failure of chemotherapy in breast cancer is caused by breast cancer stem cells (BCSCs), a minor population of cells in bulk mammary tumors. Previously, hesperetin, a citrus flavonoid, showed cytotoxicity in several cancer cells and increased cytotoxicity of doxorubicin and cisplatin. Hesperetin also inhibited osteogenic and adipocyte differentiation, however, a study of the effect of hesperetin on BCSCs has not yet been performed. Methods: In this study, we combined bioinformatics and in vitro works. A bioinformatic approach was performed to identify molecular targets, key proteins, and molecular mechanisms of hesperetin targeted at BCSCs, and genetic alterations among key genes. In addition, an in vitro study was carried out to measure the effects of hesperetin on BCSCs using the spheroids model of MCF-7 breast cancer cells (mammospheres). Results: Using a bioinformatics approach, we identified P53, PPARG, and Notch signaling as potential targets of hesperetin in inhibition of BCSCs. The in vitro study showed that hesperetin exhibits cytotoxicity on mammospheres, inhibits mammosphere and colony formation, and inhibits migration. Hesperetin modulates the cell cycle and induces apoptosis in mammospheres. Moreover, hesperetin treatment modulates the expression of p53 , PPARG , and NOTCH1 . Conclusion: Taken together, hesperetin has potential for the treatment of BCSC by targeting p53, PPARG and Notch signaling. Further investigation of the molecular mechanisms involved is required for the development of hesperetin as a BCSC-targeted drug.

Functional network analysis of p85 and PI3K as potential gene targets and mechanism of oleanolic acid in overcoming breast cancer resistance to tamoxifen

“…MDM2 encodes the MDM2 protein, which acts as a major negative regulator of the tumor suppressor p53 [ 36 ]. p53 is a barrier to cancer cell growth [ 37 ], and it is a druggable target, along with ERα [ 38 ]. These PTGs are potential targets for OA in overcoming tamoxifen resistance in breast cancer.…”

Integrated Analysis of the Expression Characteristics, Prognostic Value, and Immune Characteristics of PPARG in Breast Cancer

“…Recent studies confirmed that PPARG activation facilitates the anti-tumor effect of 6-iodolactone (Nava-Villalba et al, 2015) and hesperetin (Hermawan et al, 2021). According to a study carried by Liu stated that datasets of GEO revealed that the high expression of PPARG was associated with better prognosis in BRCA (Liu et al, 2017).…”

Chemotherapeutic potential of hesperetin for cancer treatment, with mechanistic insights: A comprehensive review

“…HSP can suppress cancer cell proliferation, viability, migration, invasion, mammosphere, and colony formation and stimulate DNA damage and apoptosis. Being a promising anticancer agent, HSP (100 μM) showed cytotoxicity on cells and prevented mammosphere, colony formation, and migration through elevating the mRNA level of p53, NOTCH1, PPARG, and reduced β-catenin resulted in apoptosis with cell cycle arrest G0/G1 phase in MCF-7 breast cancer cells [56]. Another study on a similar cell line by Choi showed that HSP (1-100 μM) administration induces apoptosis and inhibits cell proliferation after 48-72 h treatment [57].…”