Antiproliferative and Apoptotic Effects of Proteins from Black Seeds (Nigella sativa) on Human Breast MCF-7 Cancer Cell Line

Authors

Yamna Khursid, Chapman University
Basir Syed, Chapman University
Shabana U. Simjee, University of Karachi
Obaid Beg, University of Karachi
Aftab Ahmed, Chapman UniversityFollow

Document Type

Article

Publication Date

1-13-2020

Abstract

Background

Nigella sativa (NS), a member of family Ranunculaceae is commonly known as black seed or kalonji. It has been well studied for its therapeutic role in various diseases, particularly cancer. Literature is full of bioactive compounds from NS seed. However, fewer studies have been reported on the pharmacological activity of proteins. The current study was designed to evaluate the anticancer property of NS seed proteins on the MCF-7 cell line.

Methods

NS seed extract was prepared in phosphate-buffered saline (PBS), and proteins were precipitated using 80% ammonium sulfate. The crude seed proteins were partially purified using gel filtration chromatography, and peaks were resolved by SDS-PAGE. MTT assay was used to screen the crude proteins and peaks for their cytotoxic effects on MCF-7 cell line. Active Peaks (P1 and P4) were further studied for their role in modulating the expression of genes associated with apoptosis by real-time reverse transcription PCR. For protein identification, proteins were digested, separated, and analyzed with LC-MS/MS. Data analysis was performed using online Mascot, ExPASy ProtParam, and UniProt Knowledgebase (UniProtKB) gene ontology (GO) bioinformatics tools.

Results

Gel filtration chromatography separated seed proteins into seven peaks, and SDS-PAGE profile revealed the presence of multiple protein bands. Among all test samples, P1 and P4 depicted potent dose-dependent inhibitory effect on MCF-7 cells exhibiting IC₅₀ values of 14.25 ± 0.84 and 8.05 ± 0.22 μg/ml, respectively. Gene expression analysis demonstrated apoptosis as a possible cell killing mechanism. A total of 11 and 24 proteins were identified in P1 and P4, respectively. The majority of the proteins identified are located in the cytosol, associate with biological metabolic processes, and their molecular functions are binding and catalysis. Hydropathicity values were mostly in the hydrophilic range.

Conclusion

Our findings suggest NS seed proteins as a potential therapeutic agent for cancer. To our knowledge, it is the first study to report the anticancer property of NS seed proteins.

Comments

This article was originally published in BMC Complementary Medicine and Therapies, volume 20, in 2020. https://doi.org/10.1186/s12906-019-2804-1

Recommended Citation

Khurshid Y, Syed B, Simjee SU, Beg O, Ahmed A. Antiproliferative and apoptotic effects of proteins from black seeds (Nigella sativa) on human breast MCF-7 cancer cell line. BMC Complement. Med. Ther. 2020:20;5. https://doi.org/10.1186/s12906-019-2804-1

Copyright

The authors

Creative Commons License

This work is licensed under a Creative Commons Attribution 4.0 License.