Proapoptotic and Antimetastatic Properties of Supercritical CO₂ Extract of Nigella sativa Linn. Against Breast Cancer Cells

Abstract

Nigella sativa, commonly referred as black cumin, is a popular spice that has been used since the ancient Egyptians. It has traditionally been used for treatment of various human ailments ranging from fever to intestinal disturbances to cancer. This study investigated the apoptotic, antimetastatic, and anticancer activities of supercritical carbon dioxide (SC-CO2) extracts of the seeds of N. sativa Linn. against estrogen-dependent human breast cancer cells (MCF-7). Twelve extracts were prepared from N. sativa seeds using the SC-CO2 extraction method by varying pressure and temperature. Extracts were analyzed using FTIR and UV-Vis spectrometry. Cytotoxicity of the extracts was evaluated on various human cancer and normal cell lines. Of the 12 extracts, 1 extract (A3) that was prepared at 60°C and 2500 psi (*17.24 MPa) showed selective antiproliferative activity against MCF-7 cells with an IC50 of 53.34 -2.15 lg/mL. Induction of apoptosis was confirmed by evaluating caspases activities and observing the cells under a scanning electron microscope. In vitro antimetastatic properties of A3 were investigated by colony formation, cell migration, and cell invasion assays. The elevated levels of caspases in A3 treated MCF-7 cells suggest that A3 is proapoptotic. Further nuclear condensation and fragmentation studies confirmed that A3 induces cytotoxicity through the apoptosis pathway. A3 also demonstrated remarkable inhibition in migration and invasion assays of MCF-7 cells at subcytotoxic concentrations. Thus, this study highlights the therapeutic potentials of SC-CO2 extract of N. sativa in targeting breast cancer.

Use of Nigella sativa Linn. Supercritical Carbon Dioxide Extract for Targeting the Angiogenesis Cascade

“…It is suggested that phenolic content has anticancer activity [62][63][64]. These extracts showed good content of polyphenolics and total flavonoids content as published in our article [14].…”

“…The anticancer effect of these extracts was measured by MTT test on five cancerous human cell lines: MCF 7, MDA-MB-231, HCT 116, Hep G2 and PC-3, as well as one normal human cell lines: CCD-18Co. It showed that the extract A3, which was extracted at the highest temperature (60°C) and the lowest pressure (2500 psi), was the most potent extract against breast cancer (MCF 7) with IC 50 value of 53.34 ± 2.15 µg/mL as we reported previously [14]. The oil of N. sativa has showed significant anti-tumour potency [39,40].…”

“…Twelve extracts were prepared using the SFX-220 SFE system, by varying extraction pressure (2500, 3000, 4500 and 6000 psi) and temperature (32, 45 and 60°C) for 60 min dynamic according to the SC-CO 2 extraction method as described before [14]. The extracts were coded as (A1, A2, A3, B1, B2, B3, C1, C2, C3, D1, D2 and D3) and percentage yield was calculated.…”

“…Black seeds have also been used for treatment of various diseases such as fever, jaundice, inflammation and [12,13]. Recent studies have shown that N. sativa possesses medicinally active essential oils with various pharmacological activities including anti-cancer and anti-metastasis [14], anti-cestodal [15], antibacterial [16], antiviral [17], antiinflammatory [18], antinociceptive [19], and antioxidant [20] activities. N. sativa fixed oils have showed cytotoxic and immunopotentiating activities in vitro [21,22].…”

“…N. sativa fixed oils have showed cytotoxic and immunopotentiating activities in vitro [21,22]. Supercritical CO 2 extracts of N. sativa have showed potent antibacterial activities [23], also it has resulted in selective anti-proliferative and induced apoptosis of breast cancer (MCF 7) cells [14].…”