Implications of enhanced effectiveness of vincristine sulfate/ε - viniferin combination compared to vincristine sulfate only on HepG2 cells

Year 2016, Volume: 43 Issue: 4, 534 - 541, 25.12.2016

Filiz Özdemir , Zerrin İncesu Mesut Şena Nur İpek Öndera Miris Dikme

Abstract

Objective: This study was designed to investigate the
effects of ε-viniferin (ε-VNF) on the mitochondrial pathway of apoptosis
and on late apoptosis in HepG2 cell lines. To observe these effects,
ε-VNF and vincristine sulfate (VNC), anti-cancer drugs used for
treatment on HepG2 cells, were administered either alone or in
combination at different time intervals.

Methods: Mitochondrial membrane potential changes in
the cells (ΔΨm) were evaluated using cationic dye JC-1, while Bax, Bcl-2
expression levels with RT-PCR and caspase-3 activity were analyzed
using a kit. For detection of apoptotic activity, an in situ TUNEL assay
was performed.

Results: When 98.3µM ε-VNF, 52.5µM VNC and the
11.25+15.8µM VNC+ε-VNF combination were compared with the control group,
ΔΨm changes at the 6th hour were found to be 19.5%, 5.5%, 24.6%, and
3.5% , respectively. These finding show that the combination group
(24.6%) resulted in early apoptosis of the cell at the 6th hour. Bax
mRNA expression increased at the 24th hour in the VNC+ε-VNF group
compared to control group (160%), and caspase-3 activation increased in
the 1.25+15.8 µM[VNC+ε-VNF] group compared to the control group at the
48th hour. The detection of DNA fragments in HepG2 cells within 24 hours
suggests direct apoptosis.

Conclusion: These findings demonstrate that the doses
administered to the VNC+ε-VNF combination group were lower than those
administered to groups using one agent alone (e.g. VNC), the results of
which reduce the possibility of administering toxic doses.

Keywords

viniferin, vincristine, HepG2 cell

References

• 1. Ji Y, Ji C, Yue L, Xu H. Saponins isolated from Asparagus induce apoptosis in human hepatoma cell line HepG2 through a mitochondrial
• mediated pathway. Curr Oncol. 2012;19 (Suppl 2):1.
• 2. Khan M, Li T, Ahmad Khan M.K, et al. Alantolactone Induces Apoptosis in HepG2 Cells through GSH Depletion, Inhibition of STAT3 Activation, and Mitochondrial Dysfunction, Hindawi Publishing Corporation. Biomed Res Int. 2013;2013:719858.
• 3. Ohgidani M, Komizu Y, Goto K, Ueoka R. Residual powders from Shochu distillation remnants induce apoptosis in human hepatoma cells via the caspase independent pathway. J Biosci Bioeng. 2012;114:104.
• 4. Xu SP, Sun GP, Shen YX, et al. Synergistic effect of combining paeonol and cisplatin on apoptotic induction of human he patoma cell lines. Acta Pharmacol Sin. 2007;28:869.
• 5. Cao MR, Li Q, Liu ZL, et al. Harmine induces apoptosis in HepG2 cells via mitochondrial signaling pathway. Hepatobiliary Pancreat Dis Int. 2011;10:599.
• 6. Fan LL, Sun GP, Wei W, et al. Melatonin and do xorubicin synergistically induce cell apoptosis in human hepatoma cell lines. World J Gastroenterol. 2010;28:16:1473
• 7. Zghonda N, Yoshida S, Araki M, et al. Greater effectiveness of ε-viniferin in red wine than its monomer resveratrol for inhibiting vascular smooth muscle cell proliferation and migration. Biosci Biotechnol Biochem. 2011;75:1259.
• 8. Santamaria AR, Innocenti M, Mulinacci N, et al. Enhancement of viniferin production in Vitis vinifera L. cv. Alphonse Lavallée Cell uspensions by low-energy ultrasound alone and in combination with methyl jasmonate. J Agric Food Chem. 2012;60:11135.
• 9. Colin D, Lancon A, Delmas D, et al. Antiproliferative activities of resveratrol and related compounds in human hepatocyte derived HepG2 cells are associated with biochemical cell disturbance revealed by fluorescence analyses. Biochimie. 2008;90:1674.
• 10. Ozdemir F, Akalın G, Sen M, et al. Towards Novel Anti-tumor Strategies for Hepatic Cancer: ɛ-Viniferin in Combination with Vincristine Displays Pharmacodynamic Synergy at Lower Doses in HepG2 Cells. OMICS. 2014;18:324.
• 11. Yang L, Liu X, Lu Z, et al. Ursolic acid induces doxorubicin resistant HepG2 cell death via the release of apoptosis inducing factor. Cancer Lett. 2010;298:128.
• 12. Chen X, Cao Z, Zhang Y, et al. Fuzheng Qingjie Granules Inhibit Growth of Hepatoma Cells via Inducing Mitochondria Mediated poptosis and Enhancing Immune Function. Integr Cancer Ther. 2016:1–10.
• 13. Todor I.N, Lukianova N.Yu, Shvets Yu.V, et al. Metabolic changes during development of Walker-256 carcinosarcoma resistance to doxorubicin. Exp Oncol. 2015:37,1,19–22
• 14. Darzynkiewicz Z, Bedner E, Smolewski P. Flow cytometry in analysis of cell cycle and apoptosis. Semin Hematol. 2001;38:179.
• 15. Jiang CP, Ding H, Shi DH, et al. Pro-ap optotic effects of tectorigenin on human hepatocellular carcinoma HepG2 cells. World J Gastroenterol. 2012;21:18:1753.
• 16. Dikmen M, Ozturk N, Ozturk Y. The Antioxidant Potency of Punica granatum L. Fruit Peel Reduces Cell Proliferation and Induces Apoptos is on Breast Cancer. J Med Food. 2011;14:1.
• 17. Robinson L, Gallos ID, Conner SJ, et al. The effect of sperm DNA fragmentation on miscarriage rates: a systematic review and metaanalysis. Hum Reprod. 2012;27:2908-17.
• 18. Xue YQ, Di JM, Luo Y, et al. Res veratrol Oligomers for the Prevention and Treatment of Cancers. Oxid Med Cell Longev. 2014;(2014):765832.
• 19. Heiduschka G, Lill C, Seemann R, et al. The effect of resveratrol in combination with irradiation and chemotherapy: Study using Merkel cell carcinoma cell lines. Strahlenther Onkol. 2014;190:75.
• 20. Billard C, Izard J, Roman V, et al. Comparative Antiproliferative and Apoptotic Effects of Resveratrol, eviniferin and Vine-
• shots Derived Polyphenols (Vineatrols) on Chronic B Lymphocytic Leukemia Cells and Normal Human Lymphocytes. Leuk ymphoma. 2002;43:1991.
• 21. Ohguchi K, Akao Y, Matsumoto K, et al. Vaticanol Cinduced cell death is associated with inhibition of pro survival signalling in HL60 human leukemia cell line. Biosci Biotechnol Biochem. 2005;69:353.
• 22. Bhat KP, Pezzuto JM. Cancer chemopreventive activity of resveratrol. Ann N Y Acad Sci. 2002;957:210.
• 23. Jang M, Cai L, Udeani GO, et al. Cancer chemopreventive activity of resveratrol, a natura product derived from grapes. Science. 997;275(5297):218.
• 24. Baur JA, Sinclair DA. Therapeutic potential of resveratrol: the in vivo evidence. Nat Rev Drug Discov. 2006;5:493.
• 25. Gonz ́alezSarr ́ıas A, Gromek S, Niesen D, et al. Resveratrol oligomers isolated from carex species inhibit growth of humancolon tumorigenic cells mediated by cell cycle arrest. J Agric Food Chem. 2011;59:8632.
• 26. Barjot C, Tournaire M, Castagnino C, et al. Evaluation of antitumor effects of two vine stalk oligomers of resveratrol on a panel of lymphoid andmyeloid cell lines: comparison with resveratrol. Life Sci. 2007;81:1565.
• 27. Marel A.K, Lizard G, Izard J.C, et al. Inhibitory effects of transresveratrol analogsmolecules on the proliferation and the cell cycle rogression of human colon tumoral cells. Mol NutrFood Res. 2008;52:538.
• 28. Lee SM, Li ML, Tse YC, et al. Paeoniae Radix, a Chinese herbal extract, inhibit hepatoma cells growth by inducing apoptosis in a p53 independent pathway. Life Sci. 2002;27:71:2267.
• 29. Jaszewska E, Kosmider A, Kiss AK, Naruszewicz M. Oenothera paradoxa defatted seeds extract containing pentagalloylglucose and procyanidins potentiates the cytotoxicity of vincristine. J Physiol Pharmacol. 2010;61:637.
• 30. Machana S, Weerapreeyakul N, Barusrux S, et al. Cytotoxic and apoptotic effects of six herbal plants against the human hepatocarcinoma (HepG2) cell line. Chin Med. 2011;31:6:39.