Roles of p53 and Caspases in Induction of Apoptosis in MCF-7 Breast Cancer Cells Treated with a Methanolic Extract of Nigella Sativa Seeds

Abstract

Background: Nigella Sativa (NS) is an herb from the Ranunculaceae family that exhibits numerous medicinal properties and has been used as important constituent of many complementary and alternative medicines (CAMs). The ability of NS to kill cancer cells such as PC3, HeLa and hepatoma cells is well established. However, our understanding of the mode of death caused by NS remains nebulous. The objective of this study was to gain further insight into the mode and mechanism of death caused by NS in breast cancer MCF-7 cells. Materials and Methods: Human breast cancer cells (MCF-7) were treated with a methanolic extract of NS, and a dose-and time-dependent study was performed. The IC 50 was calculated using a Cell Titer Blue ® viability assay assay, and evidence for DNA fragmentation was obtained by fluorescence microscopy TUNEL assay. Gene expression was also profiled for a number of apoptosis-related genes (Caspase-3, -8, -9 and p53 genes) through qPCR. Results: The IC 50 of MCF-7 cells was 62.8µL/mL. When MCF-7 cells were exposed to 50 µL/mL and 100 µL/mL NS for 24h, 48h and 72h, microscopic examination (TUNEL assay) revealed a dose-and time-dependent increase in apoptosis. Similarly, the expression of the Caspase-3, -8, -9 and p53 genes increased significantly according to the dose and time. Conclusions: NS induced apoptosis in MCF-7 cells through both the p53 and caspase pathways. NS could potentially represent an alternative source of medicine for breast cancer therapy.

Evaluation of in vitro Anticancer Activity of Vulpinic Acid and its Apoptotic Potential Using Gene Expression and Protein Analysis

“…The study by Zu et al and our study provide similar results for alternative candidate drug molecules on the industrial area. Alhazmi et al (2014) examined molecular mechanism of NS induced apoptosis in MCF-7cell line. 29 Casp-3, Casp-8,Casp-9 and P53 gene expression changes were determined by dose-and time dependent NS.…”

“…Alhazmi et al (2014) examined molecular mechanism of NS induced apoptosis in MCF-7cell line. 29 Casp-3, Casp-8,Casp-9 and P53 gene expression changes were determined by dose-and time dependent NS. The expression levels of Casp-3, Casp-8, Casp-9 and P53 in MCF-7 cells treated with 100 μL/mL NS for 48 h markedly increased by 17.37, 4.82, 10.06 and 6.48 fold when compared with control untreated cells, respectively.…”

Effects of Polyphenol-Rich Foods on Human Health

“…In a study, a supercritical CO 2 extract of black cumin exhibited pro-apoptotic and anti-metastatic effect on MCF-7 cells in vitro [224]. Another study pointed out that the antiproliferative (IC 50 = 62.8 μL/mL) and pro-apoptotic effects of black cumin extract were through mediating both the p53 and caspase pathways [225].…”

“…Citrus fruit polysaccharides in vitro inhibiting angiogenesis and cell migration [113] Nutrients 2017, 9, 728 [141] jamun fruit NA in vitro inhibiting proliferation and inducing apoptosis [142] cranberry NA in vitro inducing apoptosis and G1 phase arrest [143] Nutrients 2017, 9, 728 [189][190][191] in vitro -inducing cell cycle arrest -lowering the invasive and metastatic potential -stimulating the expression and secretion of IFNγ [192][193][194][195][196][197] red beetroot betanin in vitro showing a dose-dependent cytotoxic effect [198] Nutrients 2017, 9, 728 inhibiting invasion by reducing MMP-9 expression via blockade of NF-κB activation [204] inhibiting invasion by suppressing invadopodium formation and MMP activity [205] inhibiting growth and sustainability of spheroid generated from adherent breast cancer cells [206] ginger 6-dehydrogingerdione in vitro inducing apoptosis and cell cycle arrest in G 2 /M phase [207] garlic diallyl disulfide in vitro inducing apoptosis though: -inhibition of histone deacetylation -inhibition of ERK and the activation of the SAPK/JNK and p38 pathways [216,217] in vitro and in vivo inhibit proliferation and metastasis via: -suppression of the SRC/Ras/ERK pathway -inactivation of the β-catenin signaling pathway [218,219] garlic diallyl trisulfide in vitro and in vivo inducing apoptosis though: -overproduction of ROS and subsequent activation of JNK and AP-1 -upregulating FAS, Bax and p53, and down-regulating Akt and Bcl-2 [220,221] in vitro -inhibiting migration and invasion; -inhibiting ER-α -decreasing protein level of FoxQ1 [213,222,223] garlic S-allyl mercaptocysteine in vitro inducing mitochondrial apoptosis and cell cycle arrest [214] garlic allicin in vitro inhibiting invasion and metastasis [215] black cumin extracts in vitro inducing apoptosis and inhibiting metastasis [224,225] black cumin thymoquinone in vitro and in vivo inducing apoptosis through: -inhibiting Akt phosphorylation -inducing p38 phosphorylation via ROS generation …”

Changes in apoptosis-related gene expression profiles in cancer cell lines exposed to usnic acid lichen secondary metabolite

“…Apoptosis that occurs through different ways can be enhanced in cells. First, candidate molecules suppress the proliferation of malignant cells, and apoptosis increases thereafter and shows a mechanistic approach to cancer chemoprevention and chemotherapy (Alhazmi et al, 2014). In an article by Singh et al published in 2013, the cell death ratio of human lung carcinoma was determined after 25-100 µM usnic acid application for 24 and 48 h. They observed that usnic acid induced mitochondrial membrane depolarization and showed approximately twofold change in apoptotic cells as regards flow cytometry analyses (Singh et al, 2013).…”