Investigation of grape and raisin extracts in induction of apoptosis and necrosis and their genetic expression on cancer stem cells (SH-SY5Y)

Jahansooz, Fatemeh; Keyhaniasl, Sara; Elahi, Shamim; Heidaripour, Ashraf

doi:10.48309/jmnc.2024.2.1

Articles in Press

Document Type : Original Article

Authors

¹ Department of Biological Science Engineering, Faculty of Modern Science and Technologies, University of Tehran, Iran

² Research Assistant of the National Organization for the Development of Brilliant Talents, Alborz, Iran

³ Department of Chemistry, Khajeh Nasir Toosi University of Technology, Tehran, Iran

10.48309/jmnc.2024.2.1

Abstract

Grapes and raisins with and without seeds were extracted using ethyl acetate/H₂O solvent. The antioxidant activity of each extract was confirmed by the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging method and free radical scavenging assay. The results confirmed the antioxidant property of the extract in such a way that the most antioxidant activity of the extract was on radical activity. Free radicals were observed in extracts of seedless raisins and seeded grapes. Their cytotoxicity was investigated on cancer stem cells (SHSY5Y) using the MTT (p-value = <0.0001) method. The results obtained from the MTT test were confirmed using the cell counting method and Trypan blue staining. Cells (SHSY5Y-1) were identified with IC50=66.87 µg/mL in seeded grapes extract and IC50=12.63 µg/mL in raisins without seeds extract. The FITC-V Annexin⁺ and PI⁺cell populations were considered as secondary apoptosis, and FITC-V Annexib^- and PI⁺ cell populations as necrosis. Data analysis was done using device software. Flow cytometry analysis showed that 62.2 % of cells underwent cell apoptosis after treatment with seedless raisin extract and 78% of cells after treatment with seeded grapes. Real time PCR method was performed for the molecular analysis of treated cells. The change was evaluated in the expression of apoptotic genes (BAX, BCL2, P53, and GAPDH). Expression of apoptotic genes for BAX and BCL2 is high under treatment with seeded grape extract. However, the expression of apoptotic genes for BCL2 and P53 is high under treatment with raisin without grape extract.

Graphical Abstract

Keywords

Main Subjects

Cancer research

References

[1]. Nirmala J.G., Narendhirakannan R., In vitro antioxidant and antimicrobial activities of grapes (Vitis vinifera L) seed and skin extracts–Muscat variety, International Journal of Pharmacy and Pharmaceutical Sciences, 2011, 3:242 [Google Scholar]

[2]. Fujimaki K., Yao G. Crack the state of silence: Tune the depth of cellular quiescence for cancer therapy, Molecular & cellular oncology, 2018, 5:e1403531 [Crossref], [Google Scholar], [Publisher]

[3]. Liu Q., Gu J., Zhang E., He L.,Yuan Z.X. Targeted delivery of therapeutics to urological cancer stem cells, Current Pharmaceutical Design, 2020, 26:2038 [Crossref], [Google Scholar], [Publisher]

[4]. Pattabiraman D.R.,Weinberg R.A. Tackling the cancer stem cells—what challenges do they pose?, Nature reviews Drug discovery, 2014, 13:497 [Crossref], [Google Scholar], [Publisher]

[5]. Miller K.D., Nogueira L., Mariotto A.B., Rowland J.H., Yabroff K.R., Alfano C.M., Jemal A., Kramer J.L., Siegel R.L. Cancer treatment and survivorship statistics, 2019, CA: a cancer journal for clinicians, 2019, 69:363 [Crossref], [Google Scholar], [Publisher]

[6]. Borah A., Raveendran S., Rochani A., Maekawa T., Kumar D. Targeting self-renewal pathways in cancer stem cells: clinical implications for cancer therapy, Oncogenesis, 2015, 4:e177 [Crossref], [Google Scholar], [Publisher]

[7]. Yuan S., Wang F., Chen G., Zhang H., Feng L., Wang L., Colman H., Keating M.J., Li X., Xu R.H. Effective elimination of cancer stem cells by a novel drug combination strategy, Stem cells, 2013, 31:23 [Crossref], [Google Scholar], [Publisher]

[8]. Stupp R. Radiotherapy plus concomitant and aduuvant temozolomide for globlastoma, The New England Journal of medicine, 2005, 352:987 [Google Scholar], [Publisher]

[9]. Grossman S.A., Batara J.F. Current management of glioblastoma multiforme, InSeminars in oncology, 2004, 31:635 [Crossref], [Google Scholar], [Publisher]

[10]. Joshi S., Guleria R., Pan J., DiPette D., Singh U. Retinoic acid receptors and tissue-transglutaminase mediate short-term effect of retinoic acid on migration and invasion of neuroblastoma SH-SY5Y cells, Oncogene, 2006, 25:240 [Crossref], [Google Scholar], [Publisher]

[11]. Pulkrabkova L., Muckova L., Hrabinova M., Sorf A., Kobrlova T., Jost P., Bezdekova D., Korabecny J., Jun D., Soukup O. Differentiated SH-SY5Y neuroblastoma cells as a model for evaluation of nerve agent-associated neurotoxicity, Archives of Toxicology, 2023, 97:2209 [Crossref], [Google Scholar], [Publisher]

[12]. Zhu M.Y., Raza M.U., Zhan Y., Fan Y. Norepinephrine upregulates the expression of tyrosine hydroxylase and protects dopaminegic neurons against 6-hydrodopamine toxicity, Neurochemistry international, 2019, 131:104549 [Crossref], [Google Scholar], [Publisher]

[13]. Saedi Z., Behbahan M. Evaluation of methanol extracts activity of seed, skin, leaf and juice from five Iranian grape cultivars on lymphocyte proliferation, Pejouhesh dar Pezeshki, 2014, 37:200 [Google Scholar], [Publisher]

[14]. Mollereau C., Zajac J.M., Roumy M. Staurosporine differentiation of NPFF2 receptor-transfected SH-SY5Y neuroblastoma cells induces selectivity of NPFF activity towards opioid receptors, Peptides, 2007, 28:1125 [Crossref], [Google Scholar], [Publisher]

[15]. Joshi S.S., Kuszynski C.A., Bagchi M., Bagchi D. Chemopreventive effects of grape seed proanthocyanidin extract on Chang liver cells, Toxicology, 2000, 155:83 [Crossref], [Google Scholar], [Publisher]

[16]. Ye X., Krohn R., Liu W., Joshi S.S., Kuszynski C., McGinn T., Bagchi M., Preuss H., Stohs S., Bagchi D. The cytotoxic effects of a novel IH636 grape seed proanthocyanidin extract on cultured human cancer cells, Stress adaptation, prophylaxis and treatment, 1999, 32:99 [Crossref], [Google Scholar], [Publisher]

[17]. Kaur M., Velmurugan B., Rajamanickam S., Agarwal R., Agarwal C. Gallic acid, an active constituent of grape seed extract, exhibits anti-proliferative, pro-apoptotic and anti-tumorigenic effects against prostate carcinoma xenograft growth in nude mice, Pharmaceutical research, 2009, 26:2133 [Crossref], [Google Scholar], [Publisher]

[18]. Aghbali A., Hosseini S.V., Delazar A., Gharavi N.K., Shahneh F.Z., Orangi M., Bandehagh A., Baradaran B. Induction of apoptosis by grape seed extract (Vitis vinifera) in oral squamous cell carcinoma, Bosnian journal of basic medical sciences, 2013, 13:186 [Crossref], [Google Scholar], [Publisher]

[19]. Erdag E. Investigation of Some Phenolic Compounds as iNOS Inhibitors: An in silico Approach, Chemical Methodologies, 2023, 7:904 [Crossref], [Publisher]

[20]. Gholamrezazadeh C., Hakimi M., Dadmehr M. A New and Safe Spirocyclic Alkoxy Phosphazene: Synthesis, Characterization, DFT, Molecular Docking and Photophysical Properties, Chemical Methodologies, 2023, 7:944 [Crossref], [Publisher]

[21]. Adyani Kalvanagh P., Adyani Kalvanagh Y. Investigating the Expression Levels of Glutathione Peroxidase and Glutathione Reductase Genes in Mastectomies Women, International Journal of Advanced Biological and Biomedical Research, 2023, 11:115 [Crossref], [Google Scholar], [Publisher]

[22]. Adyani Kalvanagh P., Adyani Kalvanagh Y. Evaluation of the effects of siRNA on Snail1 and miR-143 gene expression levels in metastatic female breast cancer cells during mastectomy, International Journal of Advanced Biological and Biomedical Research, 2023, 11:56 [Crossref], [Publisher]

[23]. Adyani Kalvanagh P., Adyani Kalvanagh Y. An Overview of Mastectomy and Its Types in the Treatment of Breast Cancer, International Journal of Advanced Biological and Biomedical Research, 2023, 11:25 [Crossref], [Publisher]

[24]. Adyani Kalvanagh P. Adyani Kalvanagh Y., Investigating the Relationship Between A/T 251 Polymorphism of IL-8 Gene and Cancer Recurrence After Lumpectomy, Eurasian Journal of Science and Technology, 2023, 3:173 [Crossref], [Publisher]

[25]. Adyani Kalvanagh P., Adyani Kalvanagh Y. Comparison of Exons 2 and 3 of DIRAS3 Gene in Mastectomies and Lumpectomies Women, Eurasian Journal of Science and Technology, 2023, 3:150 [Crossref], [Publisher]

Journal of Medicinal and Nanomaterials Chemistry

Investigation of grape and raisin extracts in induction of apoptosis and necrosis and their genetic expression on cancer stem cells (SH-SY5Y)

References

References

Articles in Press, Accepted Manuscript
Available Online from 25 February 2024

Investigation of grape and raisin extracts in induction of apoptosis and necrosis and their genetic expression on cancer stem cells (SH-SY5Y)

References

References

Articles in Press, Accepted Manuscript Available Online from 25 February 2024

Articles in Press, Accepted Manuscript
Available Online from 25 February 2024