Abstract
Purpose
To investigate the synergistic effect of resveratrol on the bystander effect of TK/GCV suicide gene system in melanoma cells.
Methods
The effect of resveratrol on the growth of B16 cells and the synergistic effect of resveratrol with or without GCV were detected by MTT assay and high content screening assay. The effect of resveratrol on GJIC function was detected by flow cytometry combined with fluorescence tracer and fluorescence microscope, and the expression of gap junction protein was detected by western blotting. Synergistic killing effect of resveratrol plus TK/GCV was tested in vivo using transplanted melanoma model.
Results
In vitro, resveratrol can enhanced GJ function and upregulated Cx32 and Cx43 protein expression in B16 cells. Resveratrol synergized with GCV to kill mixed B16 melanoma cells (20% TK+ cells and 80% TK− cells) and to improve apoptosis rate of TK− cells (the bystander effect of TK system), and the synergistic action was reversed by the GJ inhibitor AGA. In vivo, when B16 cells were mixed with 30% TK+ B16 cells, significantly reduced tumor weight and volume were observed after combinational treatment with resveratrol plus GCV as compared with GCV or resveratrol treatment alone.
Conclusions
Resveratrol could synergistically enhance the killing effect of TK/GCV suicide gene system in melanoma B16 cells and transplanted melanoma. It might be a promising adjuvant of TK/GCV therapy.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Aasen T, Mesnil M, Naus CC, Lampe PD, Laird DW (2016) Gap junctions and cancer: communicating for 50 years. Nat Rev Cancer 16:775–788. https://doi.org/10.1038/nrc.2016.105
Aasen T, Mesnil M, Naus CC, Lampe PD, Laird DW (2017) Gap junctions and cancer: communicating for 50 years. Nat Rev Cancer 17:74. https://doi.org/10.1038/nrc.2016.142
Ammerpohl O, Thormeyer D, Khan Z, Appelskog IB, Gojkovic Z, Almqvist PM, Ekstrom TJ (2004) HDACi phenylbutyrate increases bystander killing of HSV-tk transfected glioma cells. Biochem Biophys Res Commun 324:8–14. https://doi.org/10.1016/j.bbrc.2004.09.016
Bodendiek SB, Raman G (2010) Connexin modulators and their potential targets under the magnifying glass. Curr Med Chem 17:4191–4230. https://doi.org/10.2174/092986710793348563
Cottin S, Ghani K, Caruso M (2008) Bystander effect in glioblastoma cells with a predominant cytoplasmic localization of connexin43. Cancer Gene Ther 15:823–831. https://doi.org/10.1038/cgt.2008.49
Du B, Tan Y, Wu Y, Zhao P, Zhou L, Zhao Y (2004) Retroviral vector introduced in the construction of HSV1-tk/GCV antitumor suicide-gene therapeutic system. J Guangzhou Univ Trad Chin Med 21:395–398
Dyck GJB, Raj P, Zieroth S, Dyck JRB, Ezekowitz JA (2019) The effects of resveratrol in patients with cardiovascular disease and heart failure: a narrative review. Int J Mol Sci 20:904. https://doi.org/10.3390/ijms20040904
Faneca H, Duzgunes N, Pedroso de Lima MC (2019) Suicide gene therapy for oral squamous cell carcinoma. Methods Mol Biol 1895:43–55. https://doi.org/10.1007/978-1-4939-8922-5_4
Gong J, Traganos F, Darzynkiewicz Z (1994) A selective procedure for DNA extraction from apoptotic cells applicable for gel electrophoresis and flow cytometry. Anal Biochem 218:314–319. https://doi.org/10.1006/abio.1994.1184
Guan X, Wilson S, Schlender KK, Ruch RJ (1996) Gap-junction disassembly and connexin 43 dephosphorylation induced by 18 beta-glycyrrhetinic acid. Mol Carcinog 16:157–164. https://doi.org/10.1002/(sici)1098-2744(199607)16:3%3c157:Aid-mc6%3e3.0.Co;2-e
Hou CY, Tain YL, Yu HR, Huang LT (2019) The effects of resveratrol in the treatment of metabolic syndrome. Int J Mol Sci 20:535. https://doi.org/10.3390/ijms20030535
Jin ZJ (2004) About the evaluation of drug combination. Acta Pharmacol Sin 25:146–147
Karjoo Z, Chen X, Hatefi A (2016) Progress and problems with the use of suicide genes for targeted cancer therapy. Adv Drug Deliv Rev 99:113–128. https://doi.org/10.1016/j.addr.2015.05.009
Krishan A, Cabana R (2004) Flow cytometric analysis of electronic nuclear volume and DNA content in normal mouse tissues. Cell Cycle 3:380–383
Luebker SA, Koepsell SA (2019) Diverse mechanisms of BRAF inhibitor resistance in melanoma identified in clinical and preclinical studies. Front Oncol 9:268. https://doi.org/10.3389/fonc.2019.00268
Mansour SL, Thomas KR, Capecchi MR (1988) Disruption of the proto-oncogene int-2 in mouse embryo-derived stem cells: a general strategy for targeting mutations to non-selectable genes. Nature 336:348–352. https://doi.org/10.1038/336348a0
Matono S, Tanaka T, Sueyoshi S, Yamana H, Fujita H, Shirouzu K (2003) Bystander effect in suicide gene therapy is directly proportional to the degree of gap junctional intercellular communication in esophageal cancer. Int J Oncol 23:1309–1315
Mesnil M, Yamasaki H (2000) Bystander effect in herpes simplex virus-thymidine kinase/ganciclovir cancer gene therapy: role of gap-junctional intercellular communication. Cancer Res 60:3989–3999
Navarro SA, Carrillo E, Grinan-Lison C, Martin A, Peran M, Marchal JA, Boulaiz H (2016) Cancer suicide gene therapy: a patent review. Expert Opin Ther Pat 26:1095–1104. https://doi.org/10.1080/13543776.2016.1211640
Oliveira ALB et al (2017) Resveratrol role in autoimmune disease—a mini-review. Nutrients 9:1306. https://doi.org/10.3390/nu9121306
Pelin K, Hirvonen A, Linnainmaa K (1994) Expression of cell adhesion molecules and connexins in gap junctional intercellular communication deficient human mesothelioma tumour cell lines and communication competent primary mesothelial cells. Carcinogenesis 15:2673–2675. https://doi.org/10.1093/carcin/15.11.2673
Rauf A, Imran M, Suleria HAR, Ahmad B, Peters DG, Mubarak MS (2017) A comprehensive review of the health perspectives of resveratrol. Food Funct 8:4284–4305. https://doi.org/10.1039/c7fo01300k
Rincon MY, VandenDriessche T, Chuah MK (2015) Gene therapy for cardiovascular disease: advances in vector development, targeting, and delivery for clinical translation. Cardiovasc Res 108:4–20. https://doi.org/10.1093/cvr/cvv205
Robe PA, Nguyen-Khac M, Jolois O, Rogister B, Merville MP, Bours V (2005) Dexamethasone inhibits the HSV-tk/ganciclovir bystander effect in malignant glioma cells. BMC Cancer 5:32. https://doi.org/10.1186/1471-2407-5-32
Sinha D, Sarkar N, Biswas J, Bishayee A (2016) Resveratrol for breast cancer prevention and therapy: preclinical evidence and molecular mechanisms. Semin Cancer Biol 40–41:209–232. https://doi.org/10.1016/j.semcancer.2015.11.001
Tanaka T, Yamasaki H, Mesnil M (2001) Stimulation of intercellular communication of poor-communicating cells by gap-junction-competent cells enhances the HSV-TK/GCV bystander effect in vitro. Int J Cancer 91:538–542. https://doi.org/10.1002/1097-0215(200002)9999:9999%3c:aid-ijc1080%3e3.0.co;2-z
Wu X, Li Q, Feng Y, Ji Q (2018) Antitumor research of the active ingredients from traditional Chinese medical plant polygonum cuspidatum. Evid Based Complement Altern Med 2018:2313021. https://doi.org/10.1155/2018/2313021
Yan TL, Bai LF, Zhu HL, Zhang WM, Lv PC (2017) Synthesis and biological evaluation of glycyrrhetic acid derivatives as potential VEGFR2 inhibitors. ChemMedChem 12:1087–1096. https://doi.org/10.1002/cmdc.201700271
Yousef M, Vlachogiannis IA, Tsiani E (2017) Effects of resveratrol against lung cancer: in vitro and in vivo studies. Nutrients. https://doi.org/10.3390/nu9111231
Zeng L et al (2014) Construction of recombinant HSV1-tkGFP/GCV fluorescent protein monitoring system and its inhibitory effect on murine melanoma. Trad Chin Drug Res Clin Pharmacol 25:546–551
Zhao Y, de Toledo SM, Hu G, Hei TK, Azzam EI (2014) Connexins and cyclooxygenase-2 crosstalk in the expression of radiation-induced bystander effects. Br J Cancer 111:125–131. https://doi.org/10.1038/bjc.2014.276
Zulueta A, Caretti A, Signorelli P, Ghidoni R (2015) Resveratrol: a potential challenger against gastric cancer. World J Gastroenterol 21:10636–10643. https://doi.org/10.3748/wjg.v21.i37.10636
Funding
This work was supported by grants from the National Natural Science Foundation of China (Grant Nos. 81774028, 81873146, 81903606), and Natural Science Foundation of Guangdong Province (2017A030310542).
Author information
Authors and Affiliations
Contributions
YT and BD conceived and designed the study, provided technical guidance during experiments. YC, HL, YW, GZ, JL, PQ and LS carried out in vitro experiments and analyzed experimental results. YW, XL, JL and YT carried out in vivo experiments and analyzed experimental results. YC, HL and YT wrote the paper. YT and JX reviewed and edited the manuscript. All authors read and approved the final manuscript for publication.
Corresponding authors
Ethics declarations
Conflict of interest
The authors declare no conflict of interest.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Chen, Y., Li, H., Zhang, G. et al. Synergistic inhibitory effect of resveratrol and TK/GCV therapy on melanoma cells. J Cancer Res Clin Oncol 146, 1489–1499 (2020). https://doi.org/10.1007/s00432-020-03203-z
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00432-020-03203-z