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Anti-proliferation Effects of Isorhamnetin on Lung Cancer Cells in Vitro and in Vivo

  • Li, Qiong (School of Preclinical and Forensic Medicine, Sichuan University) ;
  • Ren, Fu-Qiang (Department of Thoracic Surgery, The Third People's Hospital of Chengdu) ;
  • Yang, Chun-Lei (Life Science College, Sichuan University) ;
  • Zhou, Li-Ming (School of Preclinical and Forensic Medicine, Sichuan University) ;
  • Liu, Yan-You (School of Preclinical and Forensic Medicine, Sichuan University) ;
  • Xiao, Jing (School of Preclinical and Forensic Medicine, Sichuan University) ;
  • Zhu, Ling (School of Preclinical and Forensic Medicine, Sichuan University) ;
  • Wang, Zhen-Grong (School of Preclinical and Forensic Medicine, Sichuan University)
  • Published : 2015.04.14

Abstract

Background: Isorhamnetin (Iso), a novel and essential monomer derived from total flavones of Hippophae rhamnoides that has long been used as a traditional Chinese medicine for angina pectoris and acute myocardial infarction, has also shown a spectrum of antitumor activity. However, little is known about the mechanisms of action Iso on cancer cells. Objectives: To investigate the effects of Iso on A549 lung cancer cells and underlying mechanisms. Materials and Methods: A549 cells were treated with $10{\sim}320{\mu}g/ml$ Iso. Their morphological and cellular characteristics were assessed by light and electronic microscopy. Growth inhibition was analyzed by MTT, clonogenic and growth curve assays. Apoptotic characteristics of cells were determined by flow cytometry (FCM), DNA fragmentation, single cell gel electrophoresis (comet) assay, immunocytochemistry and terminal deoxynucleotidyl transferase nick end labeling (TUNEL). Tumor models were setup by transplanting Lewis lung carcinoma cells into C57BL/6 mice, and the weights and sizes of tumors were measured. Results: Iso markedly inhibited the growth of A549 cells with induction of apoptotic changes. Iso at $20{\mu}g/ml$, could induce A549 cell apoptosis, up-regulate the expression of apoptosis genes Bax, Caspase-3 and P53, and down-regulate the expression of Bcl-2, cyclinD1 and PCNA protein. The tumors in tumor-bearing mice treated with Iso were significantly smaller than in the control group. The results of apoptosis-related genes, PCNA, cyclinD1 and other protein expression levels of transplanted Lewis cells were the same as those of A549 cells in vitro. Conclusions: Iso, a natural single compound isolated from total flavones, has antiproliferative activity against lung cancer in vitro and in vivo. Its mechanisms of action may involve apoptosis of cells induced by down-regulation of oncogenes and up-regulation of apoptotic genes.

Keywords

Isorhamnetin;apoptosis;A549;Lewis lung carcinoma;flavones

References

  1. Adachi H, Preston G, Harvat B, Dawson MI, Jetten AM (1998). Inhibition of cell proliferation and induction of apoptosis by the Retinoid AHPN in human lung carcinoma cells. Am J Respir Cell Mol Biol, 18, 323-33. https://doi.org/10.1165/ajrcmb.18.3.2974
  2. Bhattacharyya A, Mandal D, Lahiry L, Sa G, Das T (2004). Black tea protects immunocytes from tumor-induced apoptosis by changing Bcl-2/Bax ratio. Cancer Lett, 209, 147-54. https://doi.org/10.1016/j.canlet.2003.12.025
  3. Chang JY, Chang CY, Kuo CC, et al (2004). Salvinal, a novel microtubule inhibitor isolated from Salvia miltiorrhizae Bunge (Danshen), with antimitotic activity in multidrugsensitive and -resistant human tumor cells. Mol Pharmacol, 65, 77-84. https://doi.org/10.1124/mol.65.1.77
  4. Chen JJ, Ai YX, Wang JL (2010). Chemically ubiquitylated PCNA as a probe for eukaryotic translesion DNA synthesis. Nat Chem Biol, 6, 270-2. https://doi.org/10.1038/nchembio.316
  5. Das S, Das J, Samadder A, Boujedaini N, Khuda-Bukhsh AR (2012). Exp Biol Med Apigenin-induced apoptosis in A375 and A549 cells through selective action and dysfunction of mitochondria. Exp Biol Med, 237, 1433-48. https://doi.org/10.1258/ebm.2012.012148
  6. Gamet-Payrastre L, Li P, Lumeau S, et al (2000). Sulforaphane, a naturally occurring isothiocyanate, induces cell cycle arrest and apoptosis in HT29 human colon cancer cells. Cancer Res, 60, 1426-33.
  7. Gong WY, Wu JF, Liu BJ, et al (2014). Flavonoid components in Scutellaria baicalensis inhibit nicotine-induced proliferation, metastasis and lung cancer-associated inflammation in vitro. Int J Oncol, 44, 1561-70.
  8. Huigsloot M, Tijdens RB, Van de Water B (2003). Inhibition of protein kinase c enhances anticancer agent-induced loss of anchorage-independent growth regardless of protection against apoptosis by Bcl-2. Mol Pharmacol, 64, 965-73. https://doi.org/10.1124/mol.64.4.965
  9. Jacobson MD, Weil M, Raff MC (1997). Programmed cell death in animal development. Cell, 88, 347-54. https://doi.org/10.1016/S0092-8674(00)81873-5
  10. Kim BR, Hu R, Keum YS, et al (2003). Effects of glutathione on antioxidant response element-mediated gene expression and apoptosis elicited by sulforaphane. Cancer Res, 63, 7520-5.
  11. Kuo YF, Su YZ, Tseng YH, et al (2010). Flavokawain B, a novel chalcone from alpinia pricei hayata with potent apoptotic activity: involvement of ROS and GADD153 upstream of mitochondria-dependent apoptosis in HCT116 cells. Free Radical Bio Med, 49, 214-26. https://doi.org/10.1016/j.freeradbiomed.2010.04.005
  12. Li Y, Wang PZ, Zhang HY (2008). Inhibitive effect of isorhamnetin on epidermal growth factor receptor signal pathway of human gastric carcinoma cells. Chin J Public Health, 24, 1208-9.
  13. Ling YH, Jiang JD, Holland JF, Perez-Soler R (2002). Arsenic trioxide produces polymerization of microtubules and mitotic arrest before apoptosis in human tumor cell lines. Mol Pharmacol, 62, 529-38. https://doi.org/10.1124/mol.62.3.529
  14. Liu R, Meng F, Bai H, Liu Y, Liu BW (2007). Inhibitory effect of isorhamnetin and hesperidin on the oxidation of high-density lipoproteins (HDL) induced by Cu2+. J Sichuan Un iv (Med Sci Edi), 38, 961-4.
  15. Luo R, Wang JB, Zhao L, et al (2014). Synthesis and biological evaluation of baicalein derivatives as potent antitumor agents. Bioorg Med Chem Lett, 24, 1334-8. https://doi.org/10.1016/j.bmcl.2014.01.053
  16. Miyata H, DokiY, Yamamoto H, et al (2001). Overexpression of CDC25B Overrides radiation-induced G2-M arrest and results in increased apoptosis in esophageal cancer cells. Cancer Res, 61, 3188-93.
  17. Miyoshi N, Uchida K, Osawa T, Nakamura Y (2004). A link between benzyl isothiocyanate-induced cell cycle arrest and apoptosis: involvement of mitogen-activated protein kinases in the Bcl-2 phosphorylation. Cancer Res, 64, 2134-42. https://doi.org/10.1158/0008-5472.CAN-03-2296
  18. Passagne I, Evrard A, Winum JY, et al (2003). Cytotoxicity, DNA damage, and apoptosis induced by new fotemustine analogs on human melanoma cells in relation to o6-methylguanine DNA-methyltransferase expression. J Pharmacol Exp Ther, 307, 816-23. https://doi.org/10.1124/jpet.103.051938
  19. Qin LF, Ng IOL (2002). Induction of apoptosis by cisplatin and its effect on cell cycle-related proteins and cell cycle changes in hepatoma cells. Cancer lett, 75, 27-38.
  20. Radhakrishna PG, Srivastava AS, Hassanein TI, Chauhan DP, Carrier E (2004). Induction of apoptosis in human lung cancer cells by curcumin. Cancer Lett, 208, 163-70. https://doi.org/10.1016/j.canlet.2004.01.008
  21. Salivesen GS, Dixit VM (1997). Casepases: intracellular signaling by proteolysis. Cell, 91, 443-46. https://doi.org/10.1016/S0092-8674(00)80430-4
  22. Scatena CD, Stewart ZA, Mays D, et al (1998). Mitotic phosphorylation of Bcl-2 during normal cell cycle progression and Taxol-induced growth arrest. J Biol Chem, 273, 30777-84. https://doi.org/10.1074/jbc.273.46.30777
  23. Singh RP, Agarwal C, Agarwal R (2003). Inositol hexaphosphate inhibits growth, and induces G1 arrest and apoptotic death of prostate carcinoma DU145 cells: modulation of CDKICDK-cyclin and pRb-related protein-E2F complexes. Carcinogenesis, 24, 555-63. https://doi.org/10.1093/carcin/24.3.555
  24. Singh SV, Srivastava SK, Choi S, et al (2005). Sulforaphaneinduced cell death in human prostate cancer cells is initiated by reactive oxygen species. J Biol Chem, 280, 19911-24 https://doi.org/10.1074/jbc.M412443200
  25. Stoimenov I, Helleday T (2009). PCNA on the crossroad of cancer. Biochem Soc Trans, 37, 605-13. https://doi.org/10.1042/BST0370605
  26. Takagi Y, Du J, Ma XY, Nakashima I, Nagase F (2004). Phorbol 12-myristate 13-acetate protects Jurkat cells from methylglyoxal-induced apoptosis by preventing c-Jun N-terminal kinase-mediated leakage of cytochrome c in an extracellular signal-regulated kinase-dependent manner. Mol Pharmacol, 65, 778-87. https://doi.org/10.1124/mol.65.3.778
  27. Teng D, Cheng JY, Sun J, Zhang Y, Chen WN (2008). The influence of t-PA PAI-1 and vWf that osorhamnetin for ECV304 cells. Chinese Archives of Traditional Chinese Medicine, 26, 1239-40.
  28. Uthaisang W, Reutrakul V, Krachangchaeng C, Wilairat P, Fadeel B (2004). VR-3848, a novel peptide derived from Euphobiaceae, induces mitochondria-dependent apoptosis in human leukemia cells. Cancer Lett, 208, 171-8. https://doi.org/10.1016/j.canlet.2004.01.024
  29. Vasilevskaya IA, Rakitina TV, O'Dwyer PJ (2004). Quantitative effects on c-Jun N-terminal protein kinase signaling determine synergistic interaction of cisplatin and 17-allylamino-17-demethoxygeldanamycin in colon cancer cell lines. Mol Pharmacol, 65, 235-43. https://doi.org/10.1124/mol.65.1.235
  30. Wang F, Zhang SQ, Dai ZI (1998). Studies on the action of the CM4-ABP antiK562 cancer cells by SCGE. Prog Biochem Biophys, 25, 64-7.
  31. Wang ZR, Wang L, Yang HH (2000). Effect of total flavonoids of Hippophae rhamnoides on contractile mechanics and calcium transfer in stretched myocyte. Space Medical Engineering, 13, 6-9.
  32. Yang CL, Qu Y, Wang ZR, Tao DC (2004). A inhibitory effect of isorhmnetin on telomerase activity of HeLa cells. J Sichuan Univ, 35, 198-200.
  33. Zhong, LR ,Chen, X , Wei, KM (2013). Radix tetrastigma hemsleyani flavone induces apoptosis in human lung carcinoma A549 cells by modulating the MAPK pathway. Asian Pac J Cancer Prev. 14, 5983-7 https://doi.org/10.7314/APJCP.2013.14.10.5983
  34. Zhu L, Zhou LM, Yang CL, Xiao J, Wang ZR (2004). Study on apoptosis of human A549 cells induced by isorhamntin. Chin J Antibiot, 29, 687-90.
  35. Zhu L, Wang ZR, Zhou LM, et al (2005). Effects and mechanisms of Isorhamnetin on lung carcinoma1. Space Med Med Eng, 18, 381-3.
  36. Zhu L, Zhou LM, Yang CL, et al (2006). The effect and mechanism of isorhamnetin on lung cancer. Acta Pharmacologica, 27, 69.

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