DOI QR코드

DOI QR Code

Induction of Cytotoxicity and Apoptosis in Human Gastric Cancer Cell SGC-7901 by Isovaltrate Acetoxyhydrin Isolated from Patrinia heterophylla Bunge Involves a Mitochondrial Pathway and G2/M Phase Cell Cycle Arrest

  • Yang, Bo (College of Pharmaceutical Sciences, Zhejiang Chinese Medical University) ;
  • Wang, Yi-Qi (College of Pharmaceutical Sciences, Zhejiang Chinese Medical University) ;
  • Cheng, Ru-Bin (College of Pharmaceutical Sciences, Zhejiang Chinese Medical University) ;
  • Chen, Jia-Li (College of Pharmaceutical Sciences, Zhejiang Chinese Medical University) ;
  • Chen, Jin (Huadong Medicine Company Limited) ;
  • Jia, Li-Tao (Department of Gastroenterology, Tongde Hospital of Zhejiang Province) ;
  • Zhang, Ru-Song (College of Pharmaceutical Sciences, Zhejiang Chinese Medical University)
  • Published : 2013.11.30

Abstract

Background: Our previous study demonstrated cytotoxicity of a crude extract from Patrinia heterophylla Bunge (PHEB). In the present study, we aimed to investigate the effects of isovaltrate acetoxyhydrin (IA) isolated from PHEB on the gastric cancer cell SGC-7901, in order to explore a potential treatment for gastric cancer. Methods: MTT assays were employed to determine the effects of IA on cell vitality and proliferation, with monitoring of cell morphology changes and examination of apoptosis with Annexin V-PI staining. Flow cytometry was used to assess cell cycle progression and mitochondrial membrane potential. The activity of caspase 3, 9 was evaluated by spectrophotometry, and the protein levels of Bax, Bcl2 and Cyclin B1 were analyzed with Western blotting of total proteins extracted from cultured cells. Results: The results demonstrated direct toxicity of IA towards SGC-7901 cells. Evidence of apoptosis included blebbing and chromatin condensation. Annexin V-PI assays revealed early apoptosis, involving rapid depolarization of mitochondrial membranes and activity of caspase 3, 9 signaling pathways. Western blotting showed that Bcl2 and Bax proteins was down- and up-regulated, respectively, and cyclin B1 was up-regulated. Cell cycle analysis further indicated that IA could induce G2/M phase arrest in SGC-7901 cells. Conclusions: In conclusion, we believe that IA induces apoptosis of SGC-7901 cells, therefore providing a potential therapeutic agent for treatment of gastric cancer.

Keywords

References

  1. Bradford MM (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem, 72, 248-54. https://doi.org/10.1016/0003-2697(76)90527-3
  2. Chiu LC, Ho TS, Wong EY, Ooi VE (2006). Ethyl acetate extract of Patrinia scabiosaefolia downregulates anti-apoptotic Bcl-2/Bcl-X(L) expression, and induces apoptosis in human breast carcinoma MCF-7 cells independent of caspase-9 activation. J Ethnopharmacol, 105, 263-8. https://doi.org/10.1016/j.jep.2005.11.007
  3. Evans DL, Tibby M, Dive C (1994). Differential sensitivity to the induction of apoptosis by cisplatin in proliferating and quiescent immature rat thymocites is dependent of the levels of drug accumulation and DNA adduct formation. Cancer Res, 54, 1596-603.
  4. Kelly PN, Strasser A (2011). The role of Bcl-2 and its pro-survival relatives in tumourigenesis and cancer therapy. Cell Death Differ, 18, 1414-24. https://doi.org/10.1038/cdd.2011.17
  5. Kim YH, Shin KJ, Lee TG, et al (2005). G2 arrest and apoptosis by 2-amino-N-quinoline-8-ylbenzenesulfonamide (QBS), a novel cytotoxic compound. Biochem Pharmacol, 69, 1333-41. https://doi.org/10.1016/j.bcp.2004.12.019
  6. Koch U, Holzl J (1985). Uber die inhaltsstoffe von Valeriana alliariifolia. Planta Medica, 51, 172-3. https://doi.org/10.1055/s-2007-969443
  7. Lee JH, Li YC, Ip SW, et al (2008). The role of $Ca^{2+}$ in baicalein-induced apoptosis in human breast MDA-MB-231 cancer cells through mitochondria- and caspase-3- dependent pathway. Anticancer Res, 28, 1701-11.
  8. Li D, Wu LJ, Tashiro S, et al (2007). Oridonin-induced A431 cell apoptosis partially through blockage of the Ras/Raf/ERK signal pathway. J Pharmacol, 103, 56-66.
  9. Lin S, Shen YH, Li HL, et al (2009). Acylated Iridoids with Cytotoxicity from Valeriana jatamansi. J Nat Prod, 72, 650-5. https://doi.org/10.1021/np800716f
  10. Lin S, Shen YH, Zhang ZX, et al (2010). Revision of the Structures of 1,5-Dihydroxy-3,8-epoxyvalechlorine, Volvaltrate B, and Valeriotetrate C from Valeriana jatamansi and V. officinalis. J Nat Prod, 73, 1723-6. https://doi.org/10.1021/np100426j
  11. Niamh M, Pierce VK, Andrea MD, et al (2004). Nature’s sedative: Isolation and structural elucidation of valtrate from Centranthus ruber. J Chem Educ, 81, 1486-7. https://doi.org/10.1021/ed081p1486
  12. Park BY, Oh SR, Ahn KS, et al (2008). (-)-Syringaresinol inhibits proliferation of human promyelocytic HL-60 leukemia cells via G1 arrest and apoptosis. Int Immunopharmacol, 8, 967-73. https://doi.org/10.1016/j.intimp.2008.02.012
  13. Reers M, Smith TW, Chen LB (1991). J-Aggregate formation of a carbocyanine as a quantitative fluorescent indicator of membrane potential. Biochemistry, 30, 4480-6. https://doi.org/10.1021/bi00232a015
  14. Smiley ST, Reers M, Mottola-Hartshorn C, et al (1991). Intracellular heterogeneity in mitochondrial membrane potentials revealed by a J-aggregate-forming lipophilic cation JC-1. Proc Natl Acad Sci USA, 88, 3671-5. https://doi.org/10.1073/pnas.88.9.3671
  15. Szegezdi E, Macdonald DC, Ni Chonghaile T, et al (2009). Bcl-2 family on guard at the ER. Am J Physiol Cell Physiol, 296, C941-53. https://doi.org/10.1152/ajpcell.00612.2008
  16. Turner JG, Dawson J, Sullivan DM (2012). Nuclear export of proteins and drug resistance in cancer. Biochem Pharmacol, 83, 1021-32. https://doi.org/10.1016/j.bcp.2011.12.016
  17. Wahab S, Abdul A, Mohan S, et al (2009). Biological activities of pereskia bleo extracts. Int J Pharmacol, 5, 71-5. https://doi.org/10.3923/ijp.2009.71.75
  18. Watanabe K, Takatsuki H, Sonoda M, et al (2011). Anti-influenza viral effects of novel nuclear export inhibitors from Valerianae Radix and Alpinia galanga. Drug Discov Ther, 5, 26-31. https://doi.org/10.5582/ddt.v5.1.26
  19. White RJ, Reynolds IJ (1996). Mitochondrial depolarization in glutamate stimulated neurons: an early signal specific to excitotoxin exposure. J Neurosci, 16, 5688-97.
  20. Yang B, Li N, Wang YQ, et al (2011). Preliminary evaluation of antitumor effect and induction apoptosis in PC-3 cells of extract from Patrinia heterophylla Bunge. Brazilian J Pharmacognosy, 21, 471-6. https://doi.org/10.1590/S0102-695X2011005000100
  21. Yang JS, Hour MJ, Kuo SC, et al (2004). Selective induction of G2/M arrest and apoptosis in HL-60 by a potent anticancer agent, HMJ-38. Anticancer Res, 24, 1769-78.
  22. Zamzami N, Brenner C, Marzo I, et al (1998). Subcellular and submitochondrial mode of action of Bcl-2-like oncoproteins. Oncogene, 16, 2265-82. https://doi.org/10.1038/sj.onc.1201989
  23. Zamzami N, Marchetti P, Castedo M, et al (1995). Reduction in mitochondrial potential constitutes an early irreversible step of programmed lymphocyte death in vivo. J Exp Med, 181, 1661-72. https://doi.org/10.1084/jem.181.5.1661
  24. Zhang T, Li Q, Li K, et al (2008). Antitumor effects of saponin extract from Patrinia villosa (Thunb.) Juss on mice bearing U14 cervical cancer. Phytother Res, 22, 640-5. https://doi.org/10.1002/ptr.2354
  25. Zou H, Li Y, Liu X, Wang X (1999) An APAF-1-cytochrome c multimeric complex is a functional apoptosome that activates procaspase-9. J Biol Chem, 274, 11549-56. https://doi.org/10.1074/jbc.274.17.11549

Cited by

  1. /M Phase and Growth Inhibition in SGC-7901 Gastric Cancer Cells vol.15, pp.15, 2014, https://doi.org/10.7314/APJCP.2014.15.15.6437
  2. Pemetrexed is Mildly Active with Good Tolerability in Treating Patients with Gastric Cancer vol.15, pp.17, 2014, https://doi.org/10.7314/APJCP.2014.15.17.7137
  3. Mechanism of apoptosis induction in human hepatocellular carcinoma cells following treatment with a gecko peptides mixture vol.5, pp.1, 2016, https://doi.org/10.3892/br.2016.664
  4. Deacetylisovaltratum disrupts microtubule dynamics and causes G2/M-phase arrest in human gastric cancer cells in vitro vol.37, pp.12, 2016, https://doi.org/10.1038/aps.2016.91