DOI QR코드

DOI QR Code

Inductions of Caspase-, MAPK- and ROS-dependent Apoptosis and Chemotherapeutic Effects Caused by an Ethanol Extract of Scutellaria barbata D. Don in Human Gastric Adenocarcinoma Cells

  • Shim, Ji Hwan ;
  • Gim, Huijin ;
  • Lee, Soojin ;
  • Kim, Byung Joo
  • Received : 2015.02.15
  • Accepted : 2016.03.08
  • Published : 2016.06.30

Abstract

Objectives: The crude extracts of Scutellaria barbata D. Don (SB) have traditionally demonstrated inhibitory effects on numerous human cancers both in vitro and in vivo. Gastric cancer is one of the most common types of cancer on world. The authors investigated the effects of an ethanol extract of Scutellaria barbata D. Don (ESB) on the growth and survival of MKN-45 cells (a human gastric adenocarcinoma cell line). Methods: The MKN-45 cells were treated with different concentrations of ESB, and cell death was examined using an MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) assay. Analyses of sub-G1 peaks, caspase-3 and -9 activities, and mitochondrial membrane depolarizations were conducted to determine the anti-cancer effects of SB on MKN-45 cells. Also, intracellular reactive oxygen species (ROS) generation was investigated. Results: ESB inhibited the growth of MKN-45 cells, caused cell cycle arrest, and increased the sub-G1 population. In addition, ESB markedly increased mitochondrial membrane depolarization and the activities of caspase-3 and -9. ESB exerted anti-proliferative effects on MKN-45 cells by modulating the mitogen-activated protein kinase (MAPK) signaling pathway and by increasing the generation of ROS. Furthermore, combinations of anti-cancer drugs plus ESB suppressed cell growth more than treatments with an agent or ESB, and this was especially true for cisplatin, etoposide, and doxorubicin. Conclusion: ESB has a dose-dependent cytotoxic effect on MKN-45 cells and this is closely associated with the induction of apoptosis. ESB-induced apoptosis is mediated by mitochondria-, caspase- and MAPK dependent pathways. In addition, ESB enhances ROS generation and increases the chemosensitivity of MKN-45 cells. These results suggest that treatment with ESB can inhibit the proliferation and promote the apoptosis of human gastric adenocarcinoma cells by modulating the caspase-, MAPK- and ROS-dependent pathway.

Keywords

apoptosis;human gastric adenocarcinoma cell;proliferation;Scutellaria barbata D. Don

References

  1. Coccolini F, Montori G, Ceresoli M, Cima S, Valli MC, Nita GE, et al. Advanced gastric cancer: what we know and what we still have to learn. World J Gastroenterol. 2016;22(3):1139-59. https://doi.org/10.3748/wjg.v22.i3.1139
  2. Jiang Q, Li Q, Chen H, Shen A, Cai Q, Lin J, et al. Scutellaria barbata D. Don inhibits growth and induces apoptosis by suppressing IL-6-inducible STAT3 pathway activation in human colorectal cancer cells. Exp Ther Med. 2015;10(4):1602-8. https://doi.org/10.3892/etm.2015.2692
  3. Wei L, Chen Y, Lin J, Zhao J, Chen X, Xu W, et al. Scutellaria barbata D. Don induces apoptosis of human colon carcinoma cell through activation of the mitochondrion dependent pathway. J Med Plant Res. 2011;5(10):1962-70.
  4. Lin J, Chen Y, Cai Q, Wei L, Zhan Y, Shen A, et al. Scutellaria barbata D. Don inhibits colorectal cancer growth via suppression of multiple signaling pathways. Integr Cancer Ther. 2014;13(3):240-8. https://doi.org/10.1177/1534735413508811
  5. Gong T, Wang CF, Yuan JR, Li Y, Gu JF, Zhao BJ, et al. Inhibition of tumor growth and immunomodulatory effects of flavonoids and scutebarbatines of Scutellaria barbata D. Don in lewis-bearing C57BL/6 Mice. Evid Based Complement Alternat Med. 2015;2015:ID630760.
  6. Gao J, Lu WF, Dai ZJ, Lin S, Zhao Y, Li S, et al. Induction of apoptosis by total flavonoids from Scutellaria barbata D. Don in human hepatocarcinoma MHCC97-H cells via the mitochondrial pathway. Tumour Biol. 2014;35(3):2549-59. https://doi.org/10.1007/s13277-013-1336-4
  7. Xu H, Yu J, Sun Y, Xu X, Li L, Xue M, et al. Scutellaria barbata D. Don extract synergizes the antitumor effects of low dose 5-fluorouracil through induction of apoptosis and metabolism. Phytomedicine. 2013;20(10):897-903. https://doi.org/10.1016/j.phymed.2013.03.025
  8. Zhang L, Cai Q, Lin J, Fang Y, Zhan Y, Shen A, et al. Chloroform fraction of Scutellaria barbata D. Don promotes apoptosis and suppresses proliferation in human colon cancer cells. Mol Med Rep. 2014;9(2):701-6. https://doi.org/10.3892/mmr.2013.1864
  9. Majno G, Joris I. Apoptosis, oncosis, and necrosis. an overview of cell death. Am J Pathol. 1995;146(1):3-15.
  10. Saraste A, Pulkki K. Morphologic and biochemical hallmarks of apoptosis. Cardiovasc Res. 2000;45(3):528-37. https://doi.org/10.1016/S0008-6363(99)00384-3
  11. Park HS, Hong NR, Ahn TS, Kim H, Jung MH, Kim BJ. Apoptosis of AGS human gastric adenocarcinoma cells by methanolic extract of dictamnus. Pharmacogn Mag. 2015;11(S2):329-36. https://doi.org/10.4103/0973-1296.165994
  12. Nicoletti I, Migliorati G, Pagliacci MC, Grignani F, Riccardi C. A rapid and simple method for measuring thymocyte apoptosis by propidium iodide staining and flow cytometry. J Immunol Methods. 1991;139(2):271-9. https://doi.org/10.1016/0022-1759(91)90198-O
  13. Hellein KN, Kennedy EM, Harwood VJ, Gordon KV, Wang SY, Lepo JE. A filter-based propidium monoazide technique to distinguish live from membrane-compromised microorganisms using quantitative PCR. J Microbiol Methods. 2012;89(1):76-8. https://doi.org/10.1016/j.mimet.2012.01.015
  14. Wang BJ, Won SJ, Yu ZR, Su CL. Free radical scavenging and apoptotic effects of Cordyceps sinensis fractionated by supercritical carbon dioxide. Food Chem Toxicol. 2005;43(4):543-52. https://doi.org/10.1016/j.fct.2004.12.008
  15. Hong NR, Park HS, Ahn TS, Jung MH, Kim BJ. Association of a methanol extract of Rheum undulatum L. mediated cell death in AGS cells with an intrinsic apoptotic pathway. J Pharmacopuncture. 2015;18(2):26-32.
  16. Kim HS, Lee JH, Park HS, Lee GS, Kim HW, Ha KT, et al. Schizandra chinensis extracts induce apoptosis in human gastric cancer cells via JNK/p38 MAPK activation and the ROS-mediated/mitochondria-dependent pathway. Pharm Biol. 2015;53(2):212-9. https://doi.org/10.3109/13880209.2014.913297
  17. Chen BD, Ning ML, Zhou WY, Zou LL, Yu SN. Antitumour effects of Scutellaria barbata ethanol extracts in mice transplanted with human hepatocellular carcinoma (HepG2) cells. Afr J Pharm Pharmacol. 2011;5(12):1553-7. https://doi.org/10.5897/AJPP11.502
  18. Goh D, Lee YH, Ong ES. Inhibitory effects of a chemically standardized extract from Scutellaria barbata in human colon cancer cell lines, LoVo. J Agric Food Chem. 2005;53(21):8197-204. https://doi.org/10.1021/jf051506+
  19. Yin X, Zhou J, Jie C, Xing D, Zhang Y. Anticancer activity and mechanism of Scutellaria barbata extract on human lung cancer cell line A549. Life Sci. 2004;75(18):2233-44. https://doi.org/10.1016/j.lfs.2004.05.015
  20. Perez AT, Arun B, Tripathy D, Tagliaferri MA, Shaw HS, Kimmick GG, et al. A phase 1B dose escalation trial of Scutellaria barbata (BZL101) for patients with metastatic breast cancer. Breast Cancer Res Treat. 2010;120(1):111-8. https://doi.org/10.1007/s10549-009-0678-5
  21. Kim BJ. Involvement of transient receptor potential melastatin 7 channels in sophorae radix-induced apoptosis in cancer cells. J Pharmacopuncture. 2012;15(3):31-8.
  22. Hwang MW, Kim HW, Kim BJ. Involvement of transient receptor potential melastatin 7 channels in Orostachys japonicus-induced apoptosis in cancer cells. Int J Pharmacol. 2012;8(7):638-46. https://doi.org/10.3923/ijp.2012.638.646
  23. Kim BJ, Park EJ, Lee JH, Jeon JH, Kim SJ, So I. Suppression of transient receptor potential melastatin 7 channel induces cell death in gastric cancer. Cancer Sci. 2008;99(12):2502-9. https://doi.org/10.1111/j.1349-7006.2008.00982.x
  24. Lim B, Lee HJ, Kim MC, Kim BJ. Effects of ulmi pumilae cortex on AGS gastric cancer cells. J Pharmacopuncture. 2013;16(2):55-61.
  25. Lee HJ, Kim MC, Lim B, Kim BJ. Buxus microphylla var. Koreana nakai extract for the treatment of gastric cancer. J Pharmacopuncture. 2013;16(3):39-45. https://doi.org/10.3831/KPI.2013.16.016
  26. Lin WC, Lai MT, Chen HY, Ho CY, Man KM, Shen JL, et al. Protective effect of flos carthami extract against ethylene glycol‑induced urolithiasis in rats. Urol Res. 2012;40(6):655-61. https://doi.org/10.1007/s00240-012-0472-4
  27. Kim JA, Han SE, Song HJ, Chae H, Kwon YK, Kim BJ. [Effects of carthami flos on human gastric cancer cells]. Korean J Orient Physiol Pathol. 2011;25(3):466-70. Korean.

Cited by

  1. Effects of Scutellaria barbata polysaccharide on the proliferation, apoptosis and EMT of human colon cancer HT29 Cells vol.167, 2017, https://doi.org/10.1016/j.carbpol.2017.03.022
  2. Study of acetylcholinesterase activity and apoptosis in SH-SY5Y cells and mice exposed to ethanol vol.384, 2017, https://doi.org/10.1016/j.tox.2017.04.007

Acknowledgement

Supported by : National Research Foundation of Korea (NRF)