Anticancer Effect of Citrus hystrix DC. Leaf Extract and Its Bioactive Constituents Citronellol and, Citronellal on the Triple Negative Breast Cancer MDA-MB-231 Cell Line

Abstract

Triple negative breast cancer is one of the most aggressive breast cancer type with abilities of early metastasis and chemoresistance. The tropical plant Citrus hystrix DC. has been reported to promote many biological activities including anticancer. However, the effect of C. hystrix against triple negative breast cancer has not yet been identified. This study aimed to evaluate the anticancer properties of C. hystrix leaf extract and its bioactive constituents citronellol and citronellal against the triple negative breast cancer MDA-MB-231 cell line. C. hystrix leaves were powdered and sequentially macerated. The in vitro anticancer effects of C. hystrix leaf extracts, and its bioactive constituents (citronellol and citronellal) were evaluated against MDA-MB-231 cell line using cytotoxic MTT assay, cell proliferation, wound scratch migration, colony formation, cell cycle, apoptosis assay, Hoechst staining, RT-qPCR, and Western blot analysis. Results showed that crude hexane extract, citronellol, and citronellal significantly reduced cell proliferation, colony formation, and cell migration by inducing cell cycle arrest, while also inducing apoptosis in MDA-MB-231 cells through inhibition of anti-apoptotic Bcl-2 expression, leading to activation of the caspase-3-dependent pathway. This study is the first report to demonstrate the effect of C. hystrix, citronellol, and citronellal against triple negative breast cancer MDA-MB-231 cells.

Lemongrass Essential Oil Components with Antimicrobial and Anticancer Activities

“…Moreover, Limonene arrested the cell cycle in the G2/M phase and wound healing and transwell assay using Matrigel has confirmed the limonene mediated suppression of cancer cell migration and invasion [141]. Another component of the EO of lemongrass, citronellol, is also found to exert cytotoxic effects on several cancer cell lines [115,116,142]. Citronellol showed its anti-cancer activity via increased reactive oxygen species production, alterations in mitochondrial permeability, DNA fragmentation, changes in cytochrome c activities, and activation of caspase, against the MCF-7 human mammary tumor cell line [142].…”

Lemongrass Essential Oil Components with Antimicrobial and Anticancer Activities

“…Moreover, limonene arrested the cell cycle in the G2/M phase and wound healing and transwell assay using Matrigel has confirmed the limonene mediated suppression of cancer cell migration and invasion [ 140 ]. Another component of the EO of lemongrass, citronellol, is also found to exert cytotoxic effects on several cancer cell lines [ 141 , 142 , 143 ]. Citronellol showed its anticancer activity via increased reactive oxygen species production, alterations in mitochondrial permeability, DNA fragmentation, changes in cytochrome c activities, and activation of caspase, against the MCF-7 human mammary tumour cell line [ 144 ].…”

Proteomic Analysis Reveals Proteins Involved in the Mode of Action of β-Citronellol Identified From Citrus hystrix DC. Leaf Against Candida albicans

“…Due to the pleasant odor and flavor, this active compound has been used in the perfume and food industries ( Nerio et al, 2010 ; Vasconcelos et al, 2016 ). Our previous work identified citronellol from n -hexane and ethanolic crude extracts of the leaf part of C. hystrix , which demonstrated 1.42 and 4.28% relative mass, respectively ( Ho et al, 2020 ; Buakaew et al, 2021a , b ). The relative mass of an interesting citronellol was defined as its peak area divided by the total area of peaks in a single running sample.…”

“…leaf was shown by the peak region. The bioactivity of β-citronellol has been reported as having repellant ( Michaelakis et al, 2014 ), antibacterial ( Nostro et al, 2013 ), anti-inflammation ( Brito et al, 2012 ), anticancer ( Ho et al, 2020 ), anti-allergic ( Kobayashi et al, 2016 ), and antifungal properties ( Pereira et al, 2015 ; Sharma et al, 2020a ). A few reports have investigated the potential effect of β-citronellol on C. albicans ( Silva et al, 2020 ; Sharma et al, 2020b ).…”