Evaluation of in vitro anti-cancer effects of Styphnolobium japonicum root extract in human colon (HT-9), brain (U-87), and prostate (PC-3) cancer cell lines

Yıl 2020, Cilt: 50 Sayı: 2, 103 - 110, 27.08.2020

Mehmet Evren Okur Nihal Karakaş Ayşe Esra Karadağ Nurşah Öztunç İbrahim Serkut Tosyalı Fatih Demirci

Öz

Background and Aims: Styphnolobium japonicum (L.) Schott. (Sophora japonica) is a medicinal plant applied for various diseases, in the traditional medicine field. The evaluation of methanol extract of S. japonicum root derived from the Pharma Grade plant drug, was performed in terms of various in vitro biological activities. Methods: The LC-MS analysis was used for the chemical characterization of the methanol extract. The anti-cancer activity was evaluated in colon (HT-9), brain (U-87), and prostate (PC-3) cancer cells by Cell Titer Glo viability assay (Promega) and western blot analysis of PARP (Poly ADP-ribose polymerase) cleavage. Results: The relative amounts of matrine and oxymatrine in the extract were found as 0.49±0.006 mg/mL and 0.27±0.016 mg/ mL, respectively. The S. japonicum extract showed 53.17±0.97 mg of gallic acid (GA)/g corresponding to the total phenolic amounts, resulting in relatively moderate antioxidant activity (1.94±0.23 and 2.79±0.15 mg/mL) on the in vitro2,2’-azino-bis(3- ethylbenzothiazoline-6-sulfonic acid (ABTS•) and 2,2-diphenyl-1-picrylhydrazyl (DPPH•) assays. Treatment with 10 mg/mL S. japonica root extract for 24h resulted in a significant decrease in cell viability. The cell viability of U-87, HT-29, and PC-3 cancer cell lines was determined as 35±2.21%, 14±2.11%, and 46±5.67%, respectively. The extract showed 5.104, 5.012 and 0.555 mg/mL IC50 values for HT-29, U-87, and PC-3 cell lines, respectively. Particularly, the IC50 value of PC-3 cancer cell line was significantly lower than the healthy human fibroblast cells. In further, the apoptosis in S. japonicum root extract treated PC-3 cells was detected through flow cytometry analysis of Annexin V positive cells and western blot analysis of PARP cleavage. Conclusion: It can be concluded that the methanol extract in determined doses induces the apoptosis of the PC-3 cancer cells, without any significant cytotoxic effect on healthy human fibroblast cells. In addition, the LCMS analysis showed the presence of matrine and oxymatrine, which are known for their anticancer activity. To the best of our knowledge, these are the first preliminary results indicating the possible use of S. japonicum root extract. Thus, the methanol extract can be further studied for its therapeutic potential of primarily prostate and other cancer types.

Anahtar Kelimeler

Cytotoxicity, Styphnolobium japonicum (Sophora japonica), antioxidant, cancer cell lines, western blotting, flow cytometry analysis

Kaynakça

• Abdelhady, M. I. S., Kamal, A. M., Othman, S. M., Mubarak, M. S., & Hadda, T. B. (2015). Total polyphenolic content, antioxidant, cytotoxic, antidiabetic activities, and polyphenolic compounds of Sophora japonica grown in Egypt. Medicinal Chemistry Research, 24(2), 482–495.
• Blois, M. S. (1958). Antioxidant determinations by the use of a stable free radical. Nature, 181(4617), 1199–1200.
• Boulares, A. H., Yakovlev, A. G., Ivanova, V., Stoica, B. A., Wang, G., Iyer, S., & Smulson, M. (1999). Role of Poly(ADP-ribose) Polymerase (PARP) Cleavage in Apoptosis. Journal of Biological Chemistry, 274(33), 22932–22940.
• Chang, C., Liu, S-P., Fang, C-H., He, R-S., Wang, Z., Zhu, Y-Q., & JI, S-W. (2013). Effects of matrine on the proliferation of HT29 human colon cancer cells and its antitumor mechanism. Oncology Letters, 6(3), 699–704.
• Coussens, N. P., Sittampalam, G. S., Guha, R., Brimacombe, K., Grossman, A., Chung, T. D. Y., Weidner, J. R. … Austin, C. P. (2018). Assay Guidance Manual: Quantitative Biology and Pharmacology in Preclinical Drug Discovery. Clinical and Translational Science, 11(5), 461–470.
• Cragg, G. M., & Newman, D. J. (2005). Plants as a source of anticancer agents. Journal of Ethnopharmacology, 100(1–2), 72–79.
• Ding, P-L., Liao, Z-X., Huang, H., Zhou, P., & Chen, D-F. (2006). (+)-12alpha-Hydroxysophocarpine, a new quinolizidine alkaloid and related anti-HBV alkaloids from Sophora flavescens. Bioorganic & Medicinal Chemistry Letters, 16(5), 1231–5.
• Fischer, U., Janicke, R. U., & Schulze-Osthoff, K. (2003). Many cuts to ruin: A comprehensive update of caspase substrates. Cell Death and Differentiation, 10(1), 76–100.
• Hassanpour, S. H., & Dehghani, M. (2017). Review of cancer from perspective of molecular. Journal of Cancer Research and Practice, 4(4), 127–129.
• He, X., Bai, Y., Zhao, Z., Wang, X., Fang, J., Huang, L., Zeng, M., … Zheng, X. (2016). Local and traditional uses, phytochemistry, and pharmacology of Sophora japonica L.: A review. Journal of Ethnopharmacology, 187, 160–182.
• Higashiyama, K., Takeuchi, Y., Yamauchi, T., Imai, S., Kamei, J., Yajima, Y., Narita, M., & Suzuki, T. (2005). Implication of the Descending Dynorphinergic Neuron Projecting to the Spinal Cord in the (+)-Matrine- and (+)-Allomatrine-Induced Antinociceptive Effects. Biological and Pharmaceutical Bulletin, 28(5), 845–848.
• Huang, H., Wang, Q., Du, T., Lin, C., Lai, Y., Zhu, D., Wu, W., ... Li, Q. (2018). Matrine inhibits the progression of prostate cancer by promoting expression of GADD45B. Prostate, 78(5), 327–335.
• Kaufmann, S. H., Desnoyers, S., Ottaviano, Y., Davidson, N., & Poirier, G. (1993). Specific Proteolytic Cleavage of poly(ADP-ribose) Polymerase: An Early Marker of Chemotherapy-Induced Apoptosis -PubMed. Cancer Research, 53(17), 3976–3985.
• Kim, J. M., & Yun-Choi, H. S. (2008). Anti-platelet effects of flavonoids and flavonoid-glycosides from Sophora japonica. Archives of Pharmacal Research, 31(7), 886–890.
• Lee, J. W., Park, G. H., Eo, H. J., Song, H. M., Kim, M. K., Kwon, M. J., Koo, J. S., Lee, J. R., Lee, M. H., & Jeong, J. B. (2015). Anti-Cancer Activity of the Flower Bud of Sophora japonica L. through Upregulating Activating Transcription Factor 3 in Human Colorectal Cancer Cells. Korean Journal of Plant Resources, 28(3), 297–304.
• Li, Y., Li, Y., Chen, X., Liu, T., Chen, Y., He, W., Zhang, Q., & Liu, S. (2011). Autophagy is involved in anticancer effects of matrine on SGC- 7901 human gastric cancer cells. Oncology Reports, 26(1), 115–124.
• Ma, L., Wen, S., Zhan, Y., He, Y., Liu, X., & Jiang, J. (2008). Anticancer Effects of the Chinese Medicine Matrine on Murine Hepatocellular Carcinoma Cells. Planta Medica, 74(3), 245–251.
• Mihaylova, D., & Schalow, S. (2013). Antioxidant and stabilization activity of a quercetin-containing flavonoid extract obtained from Bulgarian Sophora japonica L. Brazilian Archives of Biology and Technology, 56(3), 431–438.
• Nasrollahi, S., Ghoreishi, S. M., Ebrahimabadi, A. H., & Khoobi, A. (2019). Gas chromatography-mass spectrometry analysis and antimicrobial, antioxidant and anti-cancer activities of essential oils and extracts of Stachys schtschegleevii plant as biological macromolecules. International Journal of Biological Macromolecules, 128, 718–723.
• Nobili, S., Lippi, D., Witort, E., Donnini, M., Bausi, L., Mini, E., & Capaccioli, S. (2009). Natural compounds for cancer treatment and prevention. Pharmacological Research, 59(6), 365–378.
• Okur, M. E., Ayla, Ş., Cicek Polat, D., Gunal, M. Y., Yoltaş, A., & Biceroğlu, O. (2018). Novel insight into wound healing properties of methanol extract of Capparis ovata Desf. var. palaestina Zohary fruits. Journal of Pharmacy and Pharmacology, 70(10):1401–1413.
• Pelletier, S.W. (1991). Alkaloids: Chemical and Biological Perspectives. New York, NY: Springer New York.
• Rashid, H ur., Xu, Y., Muhammad, Y., Wang, L., & Jiang, J. (2019). Research advances on anticancer activities of matrine and its derivatives: An updated overview. European Journal of Medicinal Chemistry, 161, 205–238.
• Re, R., Pellegrini, N., Proteggente, A., Pannala, A., Yang, M., & Rice- Evans, C. (1999). Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free Radical Biology and Medicine, 26(9–10), 1231–1237.
• Rodenhizer, D., Dean, T., Xu, B., Cojocari, D., & McGuigan, A.P. (2018). A three-dimensional engineered heterogeneous tumor model for assessing cellular environment and response. Nature Protocols, 13(9), 1917–1957.
• Shi, Y., Shen, G., Fang, H., Xu, C., & Hu, S. (2015). Method for quantitative determination of matrine in Sophora alopecuroides L. and its inhibitory effect on breast cancer MCF-7 cell proliferation. Biomedical Research, 26(3).
• Spanos, G. A., & Wrolstad, R. E. (1990). Influence of processing and storage on the phenolic composition of Thompson Seedless grape juice. Journal of Agricultural and Food Chemistry, 38(7), 1565–1571.
• Tang, Y-P., Li, Y-F., Hu, J., & Lou, F-C. (2002). Isolation and identification of antioxidants from Sophora japonica. Journal of Asian Natural Products Research, 4(2), 123–128.
• Tewari, M., Quan, L. T., O’Rourke, K., Desnoyers, S., Zeng, Z., Beidler, D. R., Poirier, G. G., Salvesen, G. S., & Dixit, V. M. (1995). Yama/ CPP32β, a mammalian homolog of CED-3, is a CrmA-inhibitable protease that cleaves the death substrate poly(ADP-ribose) polymerase. Cell, 81(5), 801–809.
• Ting, D., Ruwei, W., Guoyong, Z., Meizhen, C., & Songhua, L. (2002). The manufacture and clinical application of compound matrine. Acta Chinese Medicine and Pharmacology, 30(2), 47–48.
• Tomani, J. C. D., Gainkam, L. O. T., Nshutiyayesu, S., Mukazayire, M. J., Ribeiro, S. O., Stevigny, C., Frederich, M., Muganga, R., & Souopgui, J. (2018). An ethnobotanical survey and inhibitory effects on NLRP3 inflammasomes/ Caspase-1 of herbal recipes’ extracts traditionally used in Rwanda for asthma treatment. Journal of Ethnopharmacology, 227, 29–40.
• Verma, A. K., & Singh, S. (2020). Phytochemical analysis and in vitro cytostatic potential of ethnopharmacological important medicinal plants. Toxicology Reports, 7, 443–452.
• Wu, Y., Chen, J., & Cheng, Y. (2005). A sensitive and specific HPLCMS method for the determination of sophoridine, sophocarpine and matrine in rabbit plasma. Analytical and Bioanalytical Chemistry, 382(7), 1595–1600.
• Yu, P., Liu, Q., Liu, K., Yagasaki, K., Wu, E., & Zhang, G. (2009). Matrine suppresses breast cancer cell proliferation and invasion via VEGFAkt- NF-κB signaling. Cytotechnology, 59(3), 219–229.
• Zhang, G-L., Jiang, L., Yan, Q., Liu, R-H., & Zhang, L. (2015). Antitumor effect of matrine combined with cisplatin on rat models of cervical cancer. Asian Pacific Journal of Tropical Medicine, 8(12), 1055–1059.