Active Fractions of Black Rice Bran cv Cempo Ireng Inducing Apoptosis and S-phase Cell Cycle Arrest in T47D Breast Cancer Cells

Rarastoeti Pratiwi; Alfi Rizqi Amalia; Woro Anindito Sri Tunjung; Rumiyati Rumiyati

doi:10.5614/j.math.fund.sci.2019.51.1.4

Authors

Rarastoeti Pratiwi Laboratory of Biochemistry, Faculty of Biology, Universitas Gadjah Mada, Jalan Teknika Selatan, Sekip Utara,Yogyakarta, 55281, Indonesia
Alfi Rizqi Amalia Postgraduate student, Faculty of Biology, Universitas Gadjah Mada, Jalan Teknika Selatan, Sekip Utara, Yogyakarta, 55281
Woro Anindito Sri Tunjung Laboratory of Biochemistry, Faculty of Biology, Universitas Gadjah Mada, Jalan Teknika Selatan, Sekip Utara,Yogyakarta, 55281
Rumiyati Rumiyati Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Universitas Gadjah Mada, Sekip Utara, Yogyakarta, 55281

DOI:

https://doi.org/10.5614/j.math.fund.sci.2019.51.1.4

Keywords:

apoptosis, cell cycle, Cempo Ireng, cytotoxicity, S-phase

Abstract

Indonesian black rice cv Cempo Ireng was evaluated for cancer prevention using a T47D cell line model. Methanolic extract of black rice bran (BRB) showed cytotoxicity with an IC₅₀ of 522.13 g/ml. This result was in contrast to water extract of BRB, which yielded an IC₅₀ of more than 6000 g/ml. From the methanolic extract of BRB, its 6 fractions were found from the preparative TLC performance, with Rf values ranging from 0 to 0.97. According to the MTT assay of all fractions, fraction 3 (F₃) had the lowest IC₅₀ (60.17 1.72g/ml), while fraction 2 (F₂) had an IC₅₀ of 64.3 0.61g/ml. However, F₂ showed the highest induction of apoptosis, i.e. about 75.39 0.43% compared to the T47D cells without treatment (control) (about 8.39 0.16%) and with doxorubicin treatment (about 41.30 0.08%). Furthermore, fraction 5 (F₅) not only induced apoptosis but also increased the S-phase arrest percentage (60.36 2.07%), which was significantly higher than for the other treatments. Therefore, each fraction of BRB showed different cytotoxic responses to T47D cells.

References

Kristamtini, Taryono, Basunanda, P. & Murti, R.H., Genetic Variability of Rice Pericarp Color Parameters and Total Anthocyanine Content of Eleven Local Black Rice and Their Correlation, Ilmu Pertanian, 17(1), pp. 9-103, 2014.

Deng, G.F., Xu, X.R., Zhang Y., Li, D., Gan R.Y. & Li, H.B., Phenolic Compounds and Bioactivities of Pigmented Rice, Critical Reviews in Food Science and Nutrition, 53(3), pp. 296-306, 2013.

Siegel, R., Desantis, C. & Jemal, A., Breast Cancer Facts and Figures 2013-2014, American Cancer Society, Atlanta, 2013.

Holliday, D.L. & Speirs, V., Choosing The Right Cell Line for Breast Cancer Research, Breast Cancer Research, 13(4), pp. 215-221, 2011.

Rondon-Lagos, M., Cantogno, L.V.D., Marchi, C., Rangel, N., Payan-Gomez, C., Gugliotta, P., Botta, C., Bussolati, G., Ramrez-Clavijo, S.R., Pasini, B. & Sapino, A., Differences and Homologies of Chromosomal Alterations within and between Breast Cancer Cell Lines: A Clustering Analysis, Molecular Cytogenetics, 7(1), pp. 8, 2014.

Lim, L.Y., Vidnovic, N., Ellisen, L.W. & Leong, C-O., Mutant p53 Mediates Survival of Breast Cancer Cells, British Journal of Cancer, 101(9), pp. 1606-1612.

Murphy, K., Paul, T. & Mark, W., Janeway's Immunobiology, 7th edition. USA: Garland Science, 2009.

Henderson, A.J., Ollila, C.A., Kumar, A., Borresen, E.C., Raina, K., Agarwal, R. & Ryan, E.P., Chemopreventive Properties of Dietary Rice Bran: Current Status and Future Prospects, Advances in Nutrition, 3(5), pp. 643-53, 2012.

Tan, B.L., Norhaizan, M.E., Suhaniza, H.J., Lai, C.C., Norazalina, S. & Roselina, K., Antioxidant Properties and Antiproliferative Effect of Brewers' Rice Extract (Temukut) on Selected Cancer Cell Lines, International Food Research Journal, 20(5), pp. 2117-2124, 2013.

Hui, C., Bin, Y., Xiaoping, Y., Loang, Y., Chunye, C., & Mantian, M., Anticancer Activities of an Anthocyanin-Rich Extract from Black Rice Against Breast Cancer Cells In Vitro and In Vivo, Nutrition and Cancer, 62(8), pp. 1128-1136, 2010.

Hyun, J.W. & Chung, H.S., Cyanidin and Malvidin from Oryza Sativa Cv. Heugjinjubyeo Mediate Cytotoxicity Against Human Monocytic Leukemia Cells by the Arrest of G2/M Phase and Induction of Apoptosis, Journal of Agricultural and Food Chemistry, 52(8), pp. 2213-2217, 2004.

Pratiwi, R., Tunjung, W.A.S., Rumiyati & Amalia, A.R., Apoptosis Induction in Human Cervical Cancer Cells by Pigmented Rice Bran Fractions Containing Cyanidin 3-Glucoside and Peonidin 3-Glucoside, Indonesian Journal of Biotechnology, 20(1), pp. 19-26, 2015.

Kim, C.K., Cho, M.A., Choi, Y.H., Kim, J.A, Kim, Y.H., Kim, Y.K & Park, S.H., Identification and Characterization of Seed-Specific Transcription Factors Regulating Anthocyanin Biosynthesis in Black Rice, Journal of Applied Genetics, 52(2), pp. 161-169, 2012.

Meiyanto, E., Hermawan, A., Junedi, S., Fitriasari, A. & Susidarti, R.A., Nobiletin Increased Cytotoxic Activity of Doxorubicin on Mcf-7 cells but not on T47D Cells, International Journal of Phytomedicine, 3(1), pp. 29-137, 2011.

Nurhayati, A.P.D., Pratiwi, P., Wahyuono, S., Istriyati, Purnomo, H. & Syamsu., In Vitro Test and Molecular Docking of Alkaloid Compound in Marine Sponge Cinachyrella Anomala Against T47D Cell Cycle, Journal of Marine Science: Research and Development, 5(2), pp. 1-3, 2015.

Vermes, I., Haanen, C. & Reutelingsperger, C., Flow Cytometry of Apoptotic Cell Death, Journal of Immunological Methods, 243(1-2), pp. 167-190, 2000.

Meyer, B.N., Ferrigni, N.R., Putnam, J.E., Jacobsen, L.B., Nichols, D.E., & McLaughlin, J.L., Brine Shrimp: A Convenient General Bioassay for Active Plant Constituent, Planta Medica, 45(5), pp. 31-34, 1982.

Ling, W.H., Cheng, Q.X., Ma, J. & Wang, T., Red or Black Rice Decrease Atherosclerotic Plaque and Increase Antioxidant Status in Rabbits, The Journal of Nutrition, 131(5), pp. 1421-6, 2001.

Ouyang, L., Shi, Z., Zhao, S., Wang, F-T., Zhou, T-T., Liu, B. & Bao, J-K., Programmed Cell Death Pathways in Cancer: A Review of Apoptosis, Autophagy, and Programmed Necrosis, Cell Proliferation, 45(6), pp. 487-498, 2012.

Chen, P-N., Chu, S-C., Chiou, H-L., Chiang, C-L., Yang, S-F. & Hsieh, Y-S., Cyanidin 3-Glucoside and Peonidin 3-Glucoside Inhibit Tumor Cell Growth and Induce Apoptosis in Vitro and Suppress Tumor Growth in Vivo, Nutrition and Cancer, 53(2), pp. 232-243. 2005.

Sugimoto, K., Tamayose, K., Sasaki, M., Hayashi, K. & Oshimi, K., Low Dose Doxorubicin-Induced Necrosis in Jurkat Cells and its Acceleration and Conversion to Apoptosis by Antioxidants, British Journal of Haematology, 118(1), pp. 229-238, 2002.

Schantz, M., Mohn, C., Baum, M. & Richling, E., Antioxidative Efficiency of an Anthocyanin-Rich Bilberry Extract in the Human Colon Tumor Cell Lines Caco-2 and HT-29, Journal of Berry Research, 1(1), 25-33, 2010.