Involvement of non-protein thiols, mitochondrial dysfunction, reactive oxygen species and p53 in honey-induced apoptosis
- 381 Downloads
Honey is a complex mixture of different biologically active constituents. Honey possesses anti-inflammatory, antioxidant and antitumor properties. Our chief investigation was to assess the honey induced apoptosis and its molecular mechanism in colon cancer cell growth inhibition. Honey exerted antiproliferative potential against the HCT-15 and HT-29 colon cancer cells as assessed by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) assay. Flow cytometric analysis showed the increasing accumulation of hypodiploid nuclei in the sub-G1 phase of cell cycle indicating apoptosis. Honey transduced the apoptotic signal via initial depletion of intracellular non protein thiols, consequently reducing the mitochondrial membrane potential (MMP) and increasing the reactive oxygen species (ROS) generation. An increasing earlier lipid layer break was observed in the treated cells compared to the control. Honey induced apoptosis was accompanied by up-regulating the p53 and modulating the expression of pro and anti-apoptotic proteins. Further apoptosis induction was substantiated using DNA fragmentation assay and YO-PRO-1 staining. Results showed honey as a plausible candidate for induction of apoptosis through ROS and mitochondria-dependent mechanisms in colon cancer cells. This will promote honey as a potential chemotherapeutic agent against colon cancer.
KeywordsHoney Thiol ROS Apoptosis Mitochondrial dysfunction p53 YOPRO-1 DNA fragmentation Colon cancer
Authors would like to acknowledge Mr. Sanat Dey as well as his lab members for their moral support in bringing this work in proper form. Author Saravana Kumar J acknowledges the MHRD fellowship provided by IIT kharagpur. The authors would like to acknowledge the Central Research Facility (CRF) of Indian Institute of Technology, Kharagpur for providing the facility of FACS machine. Authors would also express his sincere thanks to Mr. Samiran, person-incharge of FACS machine.
Authors don’t have any conflict of interest. All work was performed with the permission from the Institute where the authors were affiliated.
- 1.American cancer society (2009) http://www.cancer.org/docroot/cri/content/cri_2_4_1x_what_are_the_key_statistics_for_colon_and_rectum_cancer.asp accessed on 03 Aug 2009
- 2.National cancer institute (2009) http://seer.cancer.gov/statfacts/html/colorect.html accessed on 03 Aug 2009
- 4.Madhavi DV, Despande SS, Salunkhe DK (1996) Food antioxidants. Dekker, New YorkGoogle Scholar
- 5.Chinthalapally V, Dhimant D, Barbara A et al (1993) Inhibitory effect of caffeic acid esters on azoxymethane-induced biochemical changes and aberrant crypt foci formation in rat colon. Cancer Res 53:4182–4188Google Scholar
- 12.Jaganathan SK, Mahitosh M (2009) Honey constituents and its apoptotic effect in colon cancer cells. JAAS 1:29–36Google Scholar
- 18.Jaganathan SK, Mahitosh M (2009) Antiproliferative effects of honey and of its polyphenols A review. J Biomed Biotechnol; in pressGoogle Scholar
- 19.Jaganathan SK, Mandal SM, Jana SK, Das S, Mandal M (2009) Studies on the phenolic profiling, anti-oxidant and cytotoxic activity of Indian honey: in-vitro Evaluation. Nat Prod Res; in pressGoogle Scholar
- 26.Constantini PC, Chernyak BC, Petronilli V et al (1996) Modulation of the mitochondrial permeability transition pore by pyridine nucleotides and dithiol oxidation at two separate sites. J Biochem Biol 271:6746–51Google Scholar