title: Biophysical Interactions of Novel Oleic Acid Conjugate and its Anticancer Potential in HeLa Cells.
abstract: Novel conjugate 2,6-Diisopropylphenol-Oleic acid (2,6P-OLA) has shown potent anticancer activity on various cancer cell lines (Khan et al. Lipids 47:973-986, 2012). In the present study, the protein-or/ and DNA-binding property of 2,6P-OLA was evaluated that could predict its potential toxic effect, in vitro. Preferential structural stability and interaction mechanism of 2,6P-OLA to human serum albumin (HSA) and calf thymus DNA (CT-DNA) was used as model molecules, employing fluorescence spectroscopy (FS) and circular dichroism (CD) methods. The binding and apoptotic activities of conjugate were determined on bacterial recombinant DNA, pBR322 and human cancer cell line, HeLa, respectively. FS studies showed a strong conjugate binding affinity to HSA with overall binding constant of K = 7.66 (±0.03) × 10(2) M(-1). Higher concentration induced detectable changes in the CD spectrum of HSA. The conjugate complexation altered HSA secondary conformation by an increase in α-helices and decrease in β-sheets. Flourescence quenching studies with CT-DNA exhibited K = 1.215 × 10(2) L mol(-1) where 2,6P-OLA efficiently displaced the ethidium bromide (EtBr) bound DNA indicating its strong competition with EtBr for intercalation. Similarly, 2,6P-OLA was able to partially bind pBR322, resulting in decrease the intensity of EtBr gradually. The conjugate significantly reduced survival of HeLa cells. Morphological studies revealed altered cell morphology, suggesting apoptotic death of HeLa cells. Overall, our data shows that 2,6P-OLA bind well with both HSA and DNA and possessed anticancer activities.
authors: Department of Pharmaceutical Chemistry, College of Pharmacy, King Saud University, Riyadh, 11451, Kingdom of Saudi Arabia, azmatbiotech@gmail.com.
authors: Azmat Ali
authors: AA
authors: Khan
authors: Amer M
authors: AM
authors: Alanazi
authors: Mumtaz
authors: M
authors: Jabeen
authors: Mohammad Khalid
authors: MK
authors: Parvez
authors: Khalid
authors: K
authors: Pervez
authors: Rizwan
authors: R
authors: Wahab
authors: Ali Saber
authors: AS
authors: Abdelhameed
authors: Arun
authors: A
authors: Chauhan
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000737:chemistry / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D002417:Cattle; D049109:Cell Proliferation / Q000187:drug effects*; D002942:Circular Dichroism; D004247:DNA / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D019301:Oleic Acid / Q000737:chemistry* / Q000378:metabolism; D011485:Protein Binding; D012709:Serum Albumin / Q000378:metabolism*; D013050:Spectrometry, Fluorescence; D013816:Thermodynamics
chemical_list: D000970:Antineoplastic Agents; D012709:Serum Albumin; D019301:Oleic Acid; D004247:DNA; C427203:calf thymus DNA
references: J Phys Chem B. 2007 Dec 13;111(49):13851-7
references: 18020438
references: Biochem Biophys Res Commun. 2001 Nov 9;288(4):954-60
references: 11689002
references: Breast Cancer Res. 2005;7(5):R645-54
references: 16168109
references: Adv Protein Chem. 1985;37:161-245
references: 3904348
references: Nanomedicine (Lond). 2013 Aug;8(8):1281-94
references: 23311988
references: J Am Chem Soc. 2006 Dec 20;128(50):16206-12
references: 17165773
references: Eur J Med Chem. 2011 Oct;46(10):4878-86
references: 21868138
references: J Inorg Biochem. 2005 Jun;99(6):1369-75
references: 15869800
references: Anal Biochem. 2006 May 15;352(2):231-42
references: 16584704
references: J Am Chem Soc. 2004 Dec 29;126(51):16686-7
references: 15612682
references: Oncol Rep. 2004 Jun;11(6):1241-52
references: 15138562
references: Chem Biodivers. 2006 Sep;3(9):975-81
references: 17193329
references: Biomacromolecules. 2007 Mar;8(3):920-7
references: 17315924
references: Anticancer Res. 2013 Jun;33(6):2517-24
references: 23749903
references: Nature. 1992 Jul 16;358(6383):209-15
references: 1630489
references: J Am Chem Soc. 2002 Nov 6;124(44):13008-17
references: 12405827
references: J Immunol Methods. 1983 Dec 16;65(1-2):55-63
references: 6606682
references: Anal Sci. 2002 Apr;18(4):391-5
references: 11999510
references: Lipids. 2012 Oct;47(10):973-86
references: 22923370
references: Bioorg Med Chem. 2010 Mar 1;18(5):1866-74
references: 20153203
references: J Am Chem Soc. 2001 Jun 27;123(25):5878-91
references: 11414820
references: Adv Protein Chem. 1994;45:153-203
references: 8154369
references: Bioorg Med Chem. 1995 Jun;3(6):723-8
references: 7582950