Alpha-Hederin, the Active Saponin of Nigella sativa, as an Anticancer Agent Inducing Apoptosis in the SKOV-3 Cell Line

Abstract

Alpha-hederin (α-HN), a pentacyclic triterpene saponin, has recently been identified as one of the active compounds of Nigella sativa, as a potential anticancer agent. However, no extensive studies on α-HN have been done as yet, as it was in the case of thymoquinone—the main ingredient of the N. sativa essential oil. To our knowledge, there are also no data available on how α-HN acts on the human cancer ovarian cell line SKOV-3. In this study we attempt to present the cytotoxic influence of α-HN on the SKOV-3 cell line by means of two methods: Real-Time xCELLigence and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The obtained IC50 values are 2.62 ± 0.04 μg/mL and 2.48 ± 0.32 μg/mL, respectively. An induction of apoptosis in SKOV-3 cells was confirmed by staining cellular nuclei with Hoechst 33342 dye and by flow cytometry analysis by binding annexin V to the cell membranes. We found that α-HN induces apoptosis in a dose-dependent manner. In the first stages of apoptosis, the mitochondrial membrane potential was found to decrease. Also, inactivation of anti-apoptotic protein Bcl-2 was observed, as well as the caspase-9 and then caspase-3/7 activation. In addition, the treatment of SKOV-3 cells with α-HN induced the cell cycle arrest of cancer cells in G0/G1 phase. The results of our investigations indicate that α-HN induces apoptosis in the SKOV-3 cell line and that the intrinsic mitochondrial pathway is involved in the programmed cancer cell death.

Bersaldegenin-1,3,5-orthoacetate induces caspase-independent cell death, DNA damage and cell cycle arrest in human cervical cancer HeLa cells

“…To monitor HeLa cell viability and proliferation we used the xCELLigence Real-Time Cell Analyzer Dual Plate (RTCA DP, ACEA Biosciences, San Diego, CA, USA) as described in our previous study (Adamska et al. 2019 ). Briefly, the cells were seeded in E-plates 16 (ACEA Bioscences) at densities of 2 × 10 4 cells/well.…”

“…The activity of caspase-3/7 was measured according with method described previously (Adamska et al. 2019 ). Briefly, the cells were seeded in 12-well plates (1 × 10 5 cells/well) and treated with bersaldegenin-1,3,5-orthoacetate at concentrations of 0.1, 0.5, 1.0, 2.0, and 5.0 µg/mL.…”

Deciphering underlying mechanism of Sars-CoV-2 infection in humans and revealing the therapeutic potential of bioactive constituents from Nigella sativa to combat COVID19: in-silico study

“…To date, a lot of therapeutic benefits of N. sativa are known and it has been shown to be effective against a wide range of illnesses like neurologic disorders, Diabetes Mellitus, hypertension, dyslipidemia, inflammatory disorders, cancer, etc. (chronic non-infectious diseases) and bacterial, fungal, viral, and parasitic infections (infectious disease) (Yimer et al, 2019 ; Adamska et al, 2019 ). Thymoquinone (2-methyl-5-propan-2-ylcyclohexa-2, 5-diene-1, 4-dione) is a major bioactive constituent of N. sativa .…”

Autophagy Modulators in Cancer Therapy

“…Described autophagy activators undergoing clinical trials and their target points, adverse effects or selected therapeutic schemes are listed in Tables 4 and 5. [194][195][196][197] NCT01552434, NCT01065662, [224]. Studies conducted by Li et al revealed that α-HN may act through increasing the ROS concentration, consequently leading to the activation of the intrinsic apoptotic pathway [225].…”

“…This compound is the main component isolated from Hedera helix L. leaves. It is also found in Nigella sativa , Kalopanax pictus and Chenopodium quinoa plants [ 224 ]. Studies conducted by Li et al revealed that α-HN may act through increasing the ROS concentration, consequently leading to the activation of the intrinsic apoptotic pathway [ 225 ].…”