Cytotoxic and apoptosis inducing effect of some pyrano [3, 2-c] pyridine derivatives against MCF-7 Breast Cancer Cells
Abstract
Anti-cancer activities of some pyrano-pyridines have been previously reported. Herein, we investigated anti-proliferative and apoptotic effects of the novel pyrano [3, 2-c] pyridine (P.P, TPM.P, 4-CP.P and 3-NP.P) compounds against MCF-7 breast cancer cells.
The MCF-7 cells were cultured in the presence of various concentrations (20-200 μM) of the compounds for 3 days and the cell viability was determined by MTT assay. Induction of apoptosis was qualitatively assayed by acridine orange/ethidium bromide (AO/EtBr) staining, DNA fragmentation assay, as well as quantitatively by Annexin V/PI double staining and cell cycle analysis.
These compounds inhibited growth and proliferation of the MCF-7 cells in a dose- and time-dependent manner. The IC50 values of P.P, TPM.P, 4-CP.P and 3-NP.P after 24 h of exposure were calculated 100 ±5.0, 180 ±6.0, 60 ±4.0 and 140 ±5.0 μM, respectively. 4-CP.P was determined as stronger compound and was chosen for further studies. The result of flow cytometric cell cycle analysis indicated an increase in sub-G1 population after 72 h treatment of the cells. Furthermore, it was accompanied with exposure of phosphatidylserine (PS) in the outer cell membrane after time course of treatment with the 4-CP.P.
Based on these observations, the pyrano [3, 2-c] pyridines can be regarded as a valuable candidate for further pharmaceutical evaluations.
Anti-cancer activities of some pyrano-pyridines have been previously reported. Herein, we investigated anti-proliferative and apoptotic effects of the novel pyrano [3, 2-c] pyridine (P.P, TPM.P, 4-CP.P and 3-NP.P) compounds against MCF-7 breast cancer cells.
The MCF-7 cells were cultured in the presence of various concentrations (20-200 μM) of the compounds for 3 days and the cell viability was determined by MTT assay. Induction of apoptosis was qualitatively assayed by acridine orange/ethidium bromide (AO/EtBr) staining, DNA fragmentation assay, as well as quantitatively by Annexin V/PI double staining and cell cycle analysis.
These compounds inhibited growth and proliferation of the MCF-7 cells in a dose- and time-dependent manner. The IC50 values of P.P, TPM.P, 4-CP.P and 3-NP.P after 24 h of exposure were calculated 100 ±5.0, 180 ±6.0, 60 ±4.0 and 140 ±5.0 μM, respectively. 4-CP.P was determined as stronger compound and was chosen for further studies. The result of flow cytometric cell cycle analysis indicated an increase in sub-G1 population after 72 h treatment of the cells. Furthermore, it was accompanied with exposure of phosphatidylserine (PS) in the outer cell membrane after time course of treatment with the 4-CP.P.
Based on these observations, the pyrano [3, 2-c] pyridines can be regarded as a valuable candidate for further pharmaceutical evaluations.
References
Aryapour H, Mahdavi M, Mohebbi SR, Zali MR, Foroumadi A (2012) Anti-proliferative and apoptotic effects of the derivatives from 4-aryl-4H-chromene family on human leukemia K562 cells. Archives of pharmacal research 35: 1573-1582.
Balalaie S, Mehrazar M, Haghighatnia Y (2013) Efficient synthesis of 2-amino-4-aryl-8-[(E)-arylmethylidene]-5, 6, 7, 8-4 H pyrano [3, 2-c] pyridine in green media. Green Chemistry Letters and Reviews 6: 101-105.
Cantrell C, Schrader K, Mamonov L, Sitpaeva G, Kustova T, Dunbar C, Wedge D (2005) Isolation and identification of antifungal and antialgal alkaloids from Haplophyllum sieversii. Journal of agricultural and food chemistry 53: 7741-7748.
Chen L, Chen J, Xu H (2013) Sasanquasaponin from Camellia oleifera Abel. induces cell cycle arrest and apoptosis in human breast cancer MCF-7 cells. Fitoterapia 84: 123-129.
Cragg GM, Newman DJ (2005) Plants as a source of anti-cancer agents. Journal of ethnopharmacology 100: 72-79.
Czyz M, Jakubowska J, Sztiller-Sikorska M (2008) STI571/doxorubicin concentration-dependent switch for diverse caspase actions in CML cell line K562. Biochemical pharmacology 75: 1761-1773.
El-Subbagh HI, Abu-Zaid SM, Mahran MA, Badria FA, Al-Obaid AM (2000) Synthesis and biological evaluation of certain α, β-unsaturated ketones and their corresponding fused pyridines as antiviral and cytotoxic agents. Journal of medicinal chemistry 43: 2915-2921.
Germano S, O'Driscoll L (2009) Breast cancer: understanding sensitivity and resistance to chemotherapy and targeted therapies to aid in personalised medicine. Current cancer drug targets 9: 398-418.
Haefner B (2003) Drugs from the deep: marine natural products as drug candidates. Drug discovery today 8: 536-544.
Hanawa F, Fokialakis N, Skaltsounis A-L (2004) Photo-activated DNA binding and antimicrobial activities of furoquinoline and pyranoquinolone alkaloids from rutaceae. Planta medica 70: 531-535.
Hassan A, Sarg M, Said M, El-Sebaey S (2013) Utility of thieno [2, 3-b] pyridine derivatives in the synthesis of some condensed heterocyclic compounds with expected biological activity. Universal Organic Chemistry 1: 2.
Holliday DL, Speirs V (2011) Choosing the right cell line for breast cancer research. Breast Cancer Res 13: 215.
Hu H, Ahn NS, Yang X, Lee YS, Kang KS (2002) Ganoderma lucidum extract induces cell cycle arrest and apoptosis in MCF‐7 human breast cancer cell. International Journal of Cancer 102: 250-253.
Huang K, Huang Y, Diao Y (2012) Wogonin induces apoptosis and down-regulates survivin in human breast cancer MCF-7 cells by modulating PI3K–AKT pathway. International immunopharmacology 12: 334-341.
Huang W-J, Wu C-L, Lin C-W, Chi L-L, Chen P-Y, Chiu C-J, Huang C-Y, Chen C-N (2010) Marchantin A, a cyclic bis (bibenzyl ether), isolated from the liverwort Marchantia emarginata subsp. tosana induces apoptosis in human MCF-7 breast cancer cells. Cancer letters 291: 108-119.
Isaka M, Tanticharoen M, Kongsaeree P, Thebtaranonth Y (2001) Structures of Cordypyridones AD, Antimalarial N-Hydroxy-and N-Methoxy-2-pyridones from the Insect Pathogenic Fungus Cordyceps n ipponica. The Journal of organic chemistry 66: 4803-4808.
Kravchenko J, Akushevich I, Seewaldt VL, Abernethy AP, Lyerly HK (2011) Breast cancer as heterogeneous disease: contributing factors and carcinogenesis mechanisms. Breast cancer research and treatment 128: 483-493.
Kumar NV, Rajendran S (2004) One-pot Synthesis and the Biological Activities of 4-Methylpyrano [3, 2-C]-quinolin-2, 5-[6H]-diones. Asian Journal of Chemistry 16: 1911.
Lage H (2003) Drug resistance in breast cancer. Cancer Therapy 1: 81-91.
Magedov IV, Manpadi M, Evdokimov NM, Elias EM, Rozhkova E, Ogasawara MA, Bettale JD, Przheval’skii NM, Rogelj S, Kornienko A (2007) Antiproliferative and apoptosis inducing properties of pyrano [3, 2-c] pyridones accessible by a one-step multicomponent synthesis. Bioorganic & medicinal chemistry letters 17: 3872-3876.
Magedov IV, Manpadi M, Ogasawara MA, Dhawan AS, Rogelj S, Van Slambrouck S, Steelant WF, Evdokimov NM, Uglinskii PY, Elias EM (2008) Structural simplification of bioactive natural products with multicomponent synthesis. 2. Antiproliferative and antitubulin activities of pyrano [3, 2-c] pyridones and pyrano [3, 2-c] quinolones. Journal of medicinal chemistry 51: 2561-2570.
Mahdavi M, Davoodi J, Zali MR, Foroumadi A (2011) Concomitant activation of caspase-9 and down-regulation of IAP proteins as a mechanism of apoptotic death in HepG2, T47D and HCT-116 cells upon exposure to a derivative from 4-aryl-4H-chromenes family. Biomedicine & Pharmacotherapy 65: 175-182.
Rahimi R, Mahdavi M, Pejman S, Zare P, Balalaei S (2015) Inhibition of cell proliferation and induction of apoptosis in K562 human leukemia cells by the derivative (3-NpC) from dihydro-pyranochromenes family. Acta Biochimica Polonica 62: 83-88.
Riccardi C, Nicoletti I (2006) Analysis of apoptosis by propidium iodide staining and flow cytometry. Nature protocols 1: 1458-1461.
Saadat YR, Saeidi N, Vahed SZ, Barzegari A, Barar J (2015) An update to DNA ladder assay for apoptosis detection. BioImpacts: BI 5: 25.
Taylor RC, Cullen SP, Martin SJ (2008) Apoptosis: controlled demolition at the cellular level. Nature reviews Molecular cell biology 9: 231-241.
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