Pentagamaboronon-0-Sorbitol Induces Apoptosis through Elevation of Reactive Oxygen Species in Triple Negative Breast Cancer Cells

Ratna Dwi Ramadani, Rohmad Yudi Utomo, Adam Hermawan, Edy Meiyanto


Breast cancer is the most common type of cancer causing mortality for women due to metastasis. More than 50% of breast cancer patients are suffered lung metastases. One strategy to target the cancerous cell is Boron Neutron Captured Therapy (BNCT) which showed high affinity toward cancer cells and reported to have anti-proliferative as well as anti-metastatic activities. Pentagamaboronon-0 (PGB-0) is a curcumin analogue substance which had reported to exert anticancer activities against Her-2 expressing as well as triple negative breast cancer cells. Despite its great potency as BNCT agent candidate, this compound also exerted several anticancer properties. Complex formation of this substance with sorbitol was achieved to improve the solubility and maximize compound’s delivery to the target cells. This study aimed to investigate the ability of Pentagamaboronon-0-Sorbitol (PGB-0-So) to modulate cell cycle and induce apoptosis especially through the mechanisms of reactive oxygen species (ROS) modulation. The 3-(4,5-dimethylthiazzol-2yl)-2,5-diphenyltetrazolium (MTT) cytotoxicity assay of PGB-0-So against 4T1 breast cancer cell line were found to exert potential effect in dose-dependent manner with lethal concentration (IC50) values of 39 μM. The cytotoxicity of PGB-0-So complex was found to be increased considerably compared with that of PGB-0. Cell cycle modulation identified using propidium iodide (PI) staining showed cell accumulation in S phase following treatment with PGB-0-So. Apoptosis induction assay analyzed using flowcytometer with Annexin V and PI staining on its IC50 dose was found to induce programmed cell death (apoptosis). The sub-IC50 treatment of this compound was also improved the cellular ROS level which also took role in apoptosis induction. These findings suggest that PGB-0-So is potential as an anticancer agent.

Keywords: Curcumin analogue, PGB-0-So, Anticancer, 4T1 cell line, ROS modulation.

Full Text:



Aggarwal, V., Tuli, H.S., Varol, A., Thakral, F. Yerer, M.B., Sak, K., et al., 2019, Role of Reactive Oxygen Species in Cancer Progression: Molecular Mechanisms and Recent Advancements, Biomolecules, 9(11), 735. CrossRef

Amalina, N.D., Nurhayati, I.P. and Meiyanto, E., 2017, Doxorubicin Induces Lamellipodia Formation and Cell Migration, Indonesian Journal of Cancer Chemoprevention, 8(2), 61–67. CrossRef

Atsumi, T., Tonosaki, K. and Fujisawa, S., 2007, Comparative cytotoxicity and ROS generation by curcumin and tetrahydrocurcumin following visible-light irradiation or treatment with horseradish peroxidase, Anticancer Research, 27(1A), 363–371.

Chen, M.-L., Sadrieh, N. and Yu, L., 2013, Impact of Osmotically Active Excipients on Bioavailability and Bioequivalence of BCS Class III Drugs, The AAPS Journal, 15(4), 1043–1050. CrossRef

Chiarugi, P. and Cirri, P., 2003, Redox regulation of protein tyrosine phosphatases during receptor tyrosine kinase signal transduction, Trends in Biochemical Sciences, 28(9), 509–514. CrossRef

Covarrubias, L., Hernandez-Garcia, D., Schnabel, D. and Salas-Vidal, E., 2008, Function of reactive oxygen species during animal development: Passive or active?, Developmental Biology, 320(1), 1–11. CrossRef

Dash, R.P., Srinivas, N.R. and Babu, R.J., 2019, Use of sorbitol as pharmaceutical excipient in the present day formulations–issues and challenges for drug absorption and bioavailability, Drug Development and Industrial Pharmacy, 45(9), 1421–1429. CrossRef

Dawood, S., 2010, Triple-negative breast cancer: epidemiology and management options, Drugs, 70(17), 2247–2258. CrossRef

Dayem, A.A., Hossain, M.K., Lee, S.B., Kim, K., Saha, S., Yang, G-M., et al., 2017, The Role of Reactive Oxygen Species (ROS) in the Biological Activities of Metallic Nanoparticles, International Journal of Molecular Sciences, 18(1), 120. CrossRef

Doumont, G., Martoriati, A., Beekman, C., Bogaerts, S., Mee, P. J., Bureau, F., Colombo, E., Alcalay, M., Bellefroid, E., Marchesi, F., Scanziani, E., Pelicci, P. G., and Marine, J., 2005, G1 checkpoint failure and increased tumor susceptibility in mice lacking the novel p53 target Ptprv, The EMBO Journal, 24(17), 3093–3103.

Dumay, A., Rincheval, V., Trotot, P., Mignotte, B. and Vayssière, J., 2006, The superoxide dismutase inhibitor diethyldithiocarbamate has antagonistic effects on apoptosis by triggering both cytochrome c release and caspase inhibition, Free Radical Biology & Medicine, 40(8), 1377–1390.

Elmore, S., 2007, Apoptosis: A Review of Programmed Cell Death, Toxicologic pathology, 35(4), 495–516. CrossRef

Ferlay, J., Soerjomataram, I., Dikshit, R., Eser, S., Mathers, C., Rebelo, M., et al., 2015, Cancer incidence and mortality worldwide: Sources, methodsand major patterns in GLOBOCAN 2012, International Journal of Cancer, 136(5), 359–386. CrossRef

He, L., He, T., Farrar, S., Ji, L., Liu, T. and Ma, X., 2017, Antioxidants Maintain Cellular Redox Homeostasis by Elimination of Reactive Oxygen Species, Cellular Physiology and Biochemistry, 44(2), 532–553. CrossRef

Karp, G., 2008, Cell and Molecular Biology : concept and experiment, in. John-Willey and Sons Inc. Hoboken., p. 654.

Kusumastuti, R., Utomo, R.Y., Khumaira, A., Putri, H., Jenie, R.I. and Meiyanto, E., 2019, Pentagamaboronon-0 increased cytotoxicity of and inhibited metastasis induction by doxorubicin in breast cancer cells, Journal of Applied Pharmaceutical Science, 9(06), 43–51. CrossRef

Larasati, Y.A., Yoneda-Kato, N., Nakamae, I., Yokoyama, T., Meiyanto, E. and Kato, J-y., 2018, Curcumin targets multiple enzymes involved in the ROS metabolic pathway to suppress tumor cell growth, Scientific Reports, 8, 2039. CrossRef

Lee, Y.T., 1983, Breast carcinoma: pattern of metastasis at autopsy, J. Surg. Oncol., 3(23), 175–180. CrossRef

Limtrakul, P. , 2007, Curcumin as Chemosentizer, in Aggarwal, B.B., Surh, Y.-J., and Shishodia, S. (eds) The Molecular Targets and Therapeutic Uses of Curcumin in Health and Disease. Boston, MA: Springer US (Advances in Experimental Medicine and Biology), 269–300. CrossRef

Liou, G.-Y. and Storz, P., 2010, Reactive oxygen species in cancer, Free radical research, 44(5), 479-496. CrossRef

Nechifor, M.T., Neagu, T-M. and Manda, G., 2009, Reactive Oxygen Species, Cancer and Anti-Cancer Therapies, Current Chemical Biology, 3(1), 22-46. CrossRef

Meiyanto, E., Putri. D.D.P., Susidarti, R.A., Murwanti, R., Sardjiman, Fitriasari, A., et al., 2014, Curcumin and its Analogues (PGV-0 and PGV-1) Enhance Sensitivity of Resistant MCF-7 Cells to Doxorubicin through Inhibition of HER2 and NF-kB Activation, Asian Pacific Journal of Cancer Prevention, 15(1), 179–184. CrossRef

Mendes, D., Alves, C., Afonso, N., Cardoso, F., Passos-Coelho, J.L., Costa, L., et al., 2015, The benefit of HER2-targeted therapies on overall survival of patients with metastatic HER2-positive breast cancer – a systematic review, Breast Cancer Research, 17(1), 140. CrossRef

Mosmann, T., 1983, Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays, Journal of Immunological Methods, 65(1–2), 55–63. CrossRef

Notarbartolo, M., Poma, P., Perri, D., Dusonchet, L., Cervello, M., D’Alessandro, N., 2005, Antitumor effects of curcumin, alone or in combination with cisplatin or doxorubicin, on human hepatic cancer cells. Analysis of their possible relationship to changes in NF-kB activation levels and in IAP gene expression, Cancer Letters, 224(1), 53–65. CrossRef

Putri, H., Jenie, R.I., Handayani, S., Kastian, R.F. and Meiyanto, E., 2016, Combination of Potassium Pentagamavunon-0 and Doxorubicin Induces Apoptosis and Cell Cycle Arrest and Inhibits Metastasis in Breast Cancer Cells, Asian Pacific journal of cancer prevention, 17(5), 2683–2688.

Qodria, L., Hairunisa, I., Utomo, R.Y., Hermawan, A. and Meiyanto, E., 2018, Anti-metastatic Activity of Curcumin Analog Pentagamaboronon-0-Sorbitol Against HER2-overexpressed MCF-7 Breast Cancer Cells, Indonesian Journal of Cancer Chemoprevention, 9(3), 118–125. CrossRef

Ramadani, R.D., Utomo, R.Y., Hermawan, A. and Meiyanto, E., 2018, Curcumin Analog Pentagamaboronon-0-Sorbitol Inhibits Cell Migration Activity of Triple Negative Breast Cancer Cell Line, Indonesian Journal of Cancer Chemoprevention, 9(3), 126–133. CrossRef

Redig, A.J. and McAllister, S.S., 2013, Breast cancer as a systemic disease: a view of metastasis, Journal of internal medicine, 274(2), 113–126. CrossRef

Redza-Dutordoir, M. and Averill-Bates, D.A., 2016, Activation of apoptosis signalling pathways by reactive oxygen species, Biochimica et Biophysica Acta, 1863(12), 2977- 2992.

Reynolds, C.P. and Maurer, B.J., 2005, Evaluating response to antineoplastic drug combinations in tissue culture models, Methods Mol Med, 110, 173- 183.

Riccardi, C. and Nicoletti, I., 2006, Analysis of apoptosis by propidium iodide staining and flow cytometry, Nature Protocols, 1(3), 1458–1461. CrossRef

Schattenberg, J.M. and Czaja, M. J., 2014, Regulation of the effects of CYP2E1-induced oxidative stress by JNK signaling, Redox Biology, 3, 7–15. CrossRef

Simon, H.-U., Haj-Yehia, A. and Levi-Schaffer, F., 2000, Role of reactive oxygen species (ROS) in apoptosis induction, Apoptosis, 5(5), 415–418. CrossRef

Son, Y., Cheong, Y-K., Kim, N-H., Chung, H-T., Kang, D.G. and Pae, H-O., 2011, Mitogen-Activated Protein Kinases and Reactive Oxygen Species: How Can ROS Activate MAPK Pathways?, Journal of Signal Transduction, 2011, 792639. CrossRef

Susnow, N., Zheng, L., Margineantu, D., and Hockenbery, D.M., 2009. Bcl-2 family proteins as regulators of oxidative stress, Semin Cancer Biol., 19(1), 42–49.

Torres, M. and Forman, H.J., 2003, Redox signaling and the MAP kinase pathways, BioFactors, 17(1–4), 287–296. CrossRef

Tsang, W.P., Chau, S.P.Y., Kong, S.K., Fung, K.P. and Kwok, T.T., 2003, Reactive oxygen species mediate doxorubicin induced p53-independent apoptosis, Life Sciences, 73,(16), 2047–2058. CrossRef

Utomo, R.Y., Putri, H., Pudjono, Susidarti, R.A., Jenie, R.I. and Meiyanto, E., 2017, Synthesis and Cytotoxic Activity of 2,5-BIS(4-Boronic Acid)Benzylidine Cyclopentanone on HER2 Overexpressed-Cancer Cells, Indonesian Journal of Pharmacy, 28(2), 74-81. CrossRef

Vermes, I., Haanen, C., Steffens-Nakken, H. and Reutelingsperger, C., 1995, A novel assay for apoptosis Flow cytometric detection of phosphatidylserine expression on early apoptotic cells using fluorescein labelled Annexin V, Journal of Immunological Methods, 184(1), 39–51. CrossRef

Watanabe, T., Hattori, Y., Ohta, Y., Ishimura, M., Nakagawa, Y., Sanada, Y., et al., 2016, Comparison of the pharmacokinetics between L-BPA and L-FBPA using the same administration dose and protocol: a validation study for the theranostic approach using [18F]-L-FBPA positron emission tomography in boron neutron capture therapy, BMC cancer, 16(1), 859. CrossRef

Xiao, W., Zheng, S., Liu, P., Zou, Y., Xie, X., Yu, P., et al., 2018, Risk factors and survival outcomes in patients with breast cancer and lung metastasis: a population- based study, Cancer Medicine, 3(7), 922–930. CrossRef

Yerlikaya, A., Okur, E. and Ulukaya, E., 2012, The p53-independent induction of apoptosis in breast cancer cells in response to proteasome inhibitor bortezomib, Tumour Biology, 33(5), 1385–1392. CrossRef

Zhang, Y. and Chen, F., 2004, Reactive oxygen species (ROS), troublemakers between nuclear factor-kappaB (NF-kappaB) and c-Jun NH(2)-terminal kinase (JNK), Cancer Research, 64(6), 1902–1905. CrossRef


Copyright (c) 2021 Ratna Dwi Ramadani, Rohmad Yudi Utomo, Adam Hermawan, Edy Meiyanto

Indexed by:





Indonesian Society for Cancer Chemoprevention