Thymoquinone Effects on Cell Viability, Apoptosis and VEGF-A Gene Expression Level in AGS(CRL-1739) Cell Line

Abstract

Background: Cancer is one of the most fatal diseases across the world and it was reported that 90% of cancer fatality depends on its angiogenesis potential. Black seed or Nigella sativa L. is a medicinal plant native to southwest Asia. N. sativa has been used for medicinal purposes for centuries and predominantly has bioactive components like Thymoquinone, which is used as a candidate for anti-cancer and anti-angiogenesis drugs. Methods: Callus was induced from leaf tissue, after that alcoholic extracts were prepared from three-month-old calluses. Thymoquinone content was measured by HPLC methods. AGS cell line was cultured and treated with standard Thymoquinone and extracts from callus. Then, cell proliferation, expression of angiogenic factor (VEGF-A gene), and apoptosis test were done by MTT assay, real-time PCR and Annexin-v kit, respectively. Results: HPLC found the maximum amount of Thymoquinone in the extract of leaf calluses, which were grown in the dark. MTT assay revealed that particular doses of extracts reduced cell proliferation. Real-time and Fluorescence- Activated Cell Sorting (FACS) results demonstrated that standard Thymoquinone and callus extracts down-regulated the VEGF-A gene expression, and all three induced apoptosis in the AGS cell line. Conclusion: It has been shown that TQ has pro-apoptotic and anti-metastatic effects on stomach cancer cell line, and these properties can introduce it as an anti-cancer drug in the near future.

Standardization of Berberis aristata DC and Nigella sativa L. Using HPTLC and GCMS and Their Antineoplasia Activity in 7,12-Dimethylbenz[a]anthracene-Induced Mouse Models

“…This indistinct feature elaborates the potential role of VEGF in tumor pathogenesis with the past studies ( Torres et al, 2016 ). Thymoquinone showed ( Figure 9D ) poor positive expression of VEGF in the ductal epithelial cells, which is similar to other studies that proved that thymoquinone attenuated tumorigenic signaling, including those controlled by EGF, FGF, VEGF, TGF, and various metastatic, angiogenic, and pro-mitogenic factors ( Rashid et al, 2019 ). α-pinene ( Figure 9E ) showed moderately strong cytoplasmic positivity for VEGF in the ductal epithelial cells and β-pinene ( Figure 9F ) depicted moderately strong cytoplasmic positivity for VEGF in the ductal epithelial cells, which mean that these isomers have less affinity toward the VEGF receptors and thus are unable to cause the apoptotic action in tumor tissues.…”

Phytochemicals Targeting VEGF and VEGF-Related Multifactors as Anticancer Therapy

“…Thymoquinone, a major monoterpene isolated from this plant, has exhibited effects against several different types of cancers in a number of preclinical studies. It attenuates tumorigenic signaling, including those controlled by EGF, FGF, VEGF, TGF-β, and various metastatic, angiogenic, and pro-mitogenic factors [124].…”

Chemopreventive and Anticancer Effects of Thymoquinone: Cellular and Molecular Targets

“…TQ attenuates tumor angiogenesis by inhibiting the expression of VEGF as well as the signaling pathways stimulated by VEGF. TQ also downregulates the expression VEGF in several different cell lines derived from gastric carcinoma [129], osteosarcoma [130], colon carcinoma [131], cholangiocarcinoma [63], prostate cancer cells [95], multiple myeloma cells [101,102], and leukemia [70]. In addition to VEGF, TQ has been shown to inhibit the expression of several other angiogenic chemokines, such as CXCL5/ ENA-78 and CXCL1/Gro-alpha cytokines in non-small lung cancer and small cell lung cancer cell lines [59].…”