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Antiproliferative and apoptotic effects of proteins from black seeds (Nigella sativa) on human breast MCF-7 cancer cell line.
BMC Complement Med Ther. 2020 Jan 13; 20(1):5.BC

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.

Authors+Show Affiliations

Biomedical and Pharmaceutical Sciences, Chapman University School of Pharmacy, 9401 Jeronimo Road, Irvine, CA, 92618, USA. Dr. Panjwani Center for Molecular Medicine and Drug Research, International Center for Chemical and Biological Sciences, University of Karachi, Karachi, 75270, Pakistan.Biomedical and Pharmaceutical Sciences, Chapman University School of Pharmacy, 9401 Jeronimo Road, Irvine, CA, 92618, USA.Dr. Panjwani Center for Molecular Medicine and Drug Research, International Center for Chemical and Biological Sciences, University of Karachi, Karachi, 75270, Pakistan.Dr. Panjwani Center for Molecular Medicine and Drug Research, International Center for Chemical and Biological Sciences, University of Karachi, Karachi, 75270, Pakistan.Biomedical and Pharmaceutical Sciences, Chapman University School of Pharmacy, 9401 Jeronimo Road, Irvine, CA, 92618, USA. aahmed@chapman.edu.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

32020890

Citation

Khurshid, Yamna, et al. "Antiproliferative and Apoptotic Effects of Proteins From Black Seeds (Nigella Sativa) On Human Breast MCF-7 Cancer Cell Line." BMC Complementary Medicine and Therapies, vol. 20, no. 1, 2020, p. 5.
Khurshid Y, Syed B, Simjee SU, et al. 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(1):5.
Khurshid, Y., Syed, B., Simjee, S. U., Beg, O., & Ahmed, A. (2020). Antiproliferative and apoptotic effects of proteins from black seeds (Nigella sativa) on human breast MCF-7 cancer cell line. BMC Complementary Medicine and Therapies, 20(1), 5. https://doi.org/10.1186/s12906-019-2804-1
Khurshid Y, et al. Antiproliferative and Apoptotic Effects of Proteins From Black Seeds (Nigella Sativa) On Human Breast MCF-7 Cancer Cell Line. BMC Complement Med Ther. 2020 Jan 13;20(1):5. PubMed PMID: 32020890.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Antiproliferative and apoptotic effects of proteins from black seeds (Nigella sativa) on human breast MCF-7 cancer cell line. AU - Khurshid,Yamna, AU - Syed,Basir, AU - Simjee,Shabana U, AU - Beg,Obaid, AU - Ahmed,Aftab, Y1 - 2020/01/13/ PY - 2019/07/30/received PY - 2019/12/22/accepted PY - 2020/2/6/entrez PY - 2020/2/6/pubmed PY - 2020/7/30/medline KW - Anticancer activity KW - Apoptosis KW - LC-MS/MS KW - Nigella sativa seed KW - Protein purification SP - 5 EP - 5 JF - BMC complementary medicine and therapies JO - BMC Complement Med Ther VL - 20 IS - 1 N2 - 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. SN - 2662-7671 UR - https://www.unboundmedicine.com/medline/citation/32020890/Antiproliferative_and_apoptotic_effects_of_proteins_from_black_seeds__Nigella_sativa__on_human_breast_MCF_7_cancer_cell_line_ L2 - https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/32020890/ DB - PRIME DP - Unbound Medicine ER -