Cardanol isolated from Thai Apis mellifera propolis induces cell cycle arrest and apoptosis of BT-474 breast cancer cells via p21 upregulation

Abstract

BackgroundCardanol was previously reported to be an antiproliferative compound purified from Thai Apis mellifera propolis. By morphology, it could induce the cell death to many cancer cell lines but not the control (non-transformed human foreskin fibroblast cell line, Hs27). Here, it was aimed to evaluate the molecular effects of cardanol on breast cancer derived cell line (BT-474).MethodsMorphological changes in BT-474 cells induced by cardanol compared to doxorubicin were evaluated by light microscopy, cytotoxicity by using the 3- (4, 5-dimethyl-thiazol-2-yl) 2, 5-diphenyl-tetrazolium bromide (MTT) assay, induction of cell cycle arrest and cell death by flow cytometric analysis of propidium iodide and annexin-V stained cells, and changes in the expression level of genes involved in the control of apoptosis and the cell cycle by quantitative reverse transcriptase-PCR (qRT-PCR) and western blot analyses.ResultsIt revealed that cardanol induced a time- and dose-dependent cytotoxicity along with cell shrinkage and detachment from substratum. Cardanol caused cell cycle arrest at the G1 subphase (as opposed to at the G2/M subphase seen with doxorubicin) and cell death by late apoptosis, with both late apoptosis (27.2 ± 1.1 %) and necrosis (25.4 ± 1.4 %) being found in cardanol treated cells after 72 h, compared to a lower proportion of apoptosis (4.3 ± 0.4 %) and higher proportion of necrosis (35.8 ± 13.0 %) induced by doxorubicin. Moreover, cardanol changed the transcript expression levels of genes involved in the control of apoptosis (increased DR5 and Bcl-2 expression and decreased Mcl-1, MADD and c-FLIPP) and cell division (increased p21 and E2FI and decreased cyclin D1, cyclin E, CDK4 and CDK2 expression), as well as increasing the level of p21 p-ERK, p-JNK and p-p38 and decreasing cyclin D. This accounts for the failure to progress from the G1 to the S subphase.ConclusionCardanol is a potential chemotherapeutic agent for breast cancer.Electronic supplementary materialThe online version of this article (doi:10.1186/s40199-015-0138-1) contains supplementary material, which is available to authorized users.

Melittin Induced G1 Cell Cycle Arrest and Apoptosis in Chago-K1 Human Bronchogenic Carcinoma Cells and Inhibited the Differentiation of THP-1 Cells into Tumour- Associated Macrophages

“…Here, melittin-induced apoptosis of ChaGo-K1 was supported by the observed morphological changes (cell shrinkage, round cell formation, nucleus and organelle condensation, cell floating, decrease in viable cell density and presence of cell debris) and the significantly higher and lower expression of Bcl-2 and MADD transcripts, respectively. Similar altered gene expression was seen in apoptosis of breast cancer BT-474 cells induced by cardanol (Buahorm et al, 2015). However, whether melittin induced apoptosis or necrosis depended on the dose.…”

“…Quantitative reverse transcriptase polymerase chain reaction (RT-qPCR) was used to amplify CatS, Bcl-2, MADD and ß-actin mRNA using the One Step SYBR PrimeScript RT-qPCR Kit II (Takara, Tokyo, Japan) as per the manufacturer's protocol. Respective forward and reverse primers of CatS, including the optimal condition of RT-qPCR were obtained from , while those of ß-actin, Bcl-2 and MADD, including the optimal condition of RT-qPCR, were obtained from Buahorm et al (2015). The relative expression levels of target genes were normalized to the expression level of the ß-actin gene as control.…”

Antiproliferative Activity of Chemically Characterized Propolis from Turkey and Its Mechanisms of Action

“…In some previous studies investigating anticancer effects of main active ingredients of propolis such as caffeic acid phenyl ester (CAPE), caffeic acid (CA) or cardanol on different breast cancer cell lines revealed that these compounds can inhibit cell cycle progression [18] and activate p21. [19] In our study, propolis treatment induced p21 protein levels in addition to p53, significantly increased sub G0 phase while decreasing S and G2/M phases in MCF7 cells in addition to promoting apoptosis and decreasing viability significantly.…”

The preservative effect of Thai propolis extract on the viability of human periodontal ligament cells

“…However, no additional proliferative effect of this propolis extract on PDL cell growth was demonstrated compared with a significant increase in cell growth by PMA treatment. Interestingly, the significant inhibition of PDL cell proliferation by treatment with 10 mg ml À1 of this extract is in line with the antiproliferative effect of Thai propolis extract on several different cancer cell lines (17,19). In this study, ethanol rather than water was used to extract Thai propolis because the ethanol extract of Thai propolis contains higher flavonoid compounds (20).…”