Asian Pacific Journal of Cancer Prevention

Asian Pacific Journal of Cancer Prevention (아시아태평양암예방학회)
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Targeting of COX-2 Expression by Recombinant Adenovirus shRNA Attenuates the Malignant Biological Behavior of Breast Cancer Cells

  • Tu, Bo (Department of Ultrasonic Medicine, The First Affiliated Hospital of Chongqing Medical University) ;
  • Ma, Ting-Ting (Key Laboratory of Laboratory Medical Diagnostics of Education Ministry, Chongqing Medical University) ;
  • Peng, Xiao-Qiong (Department of Ultrasonic Medicine, The First Affiliated Hospital of Chongqing Medical University) ;
  • Wang, Qin (Key Laboratory of Laboratory Medical Diagnostics of Education Ministry, Chongqing Medical University) ;
  • Yang, Hong (Department of Ultrasonic Medicine, The First Affiliated Hospital of Chongqing Medical University) ;
  • Huang, Xiao-Ling (Department of Ultrasonic Medicine, The First Affiliated Hospital of Chongqing Medical University)
  • Published : 2014.11.06
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Abstract

Background: Cyclooxygenase-2 (COX-2), considered to have tumor-promoting potential, is highly expressed in a variety of tumors, including breast cancer. Since the functions and action mechanisms of COX-2 in breast cancer have not been fully elucidated, in the present study, the effects of target inhibiting COX-2 with recombinant adenovirus Ad-COX-2-shRNA on malignant biological behavior were investigated in representative cell lines. Materials and Methods: Breast cancer MDA-MB-231 and MCF-7 cells were transfected with Ad-COX-2-shRNA and COX-2 expression was tested by RT-PCR and Western blotting. Changes in proliferation, apoptosis and invasion of breast cancer cells were detected with various assays including MTT, colony forming, flowcytometry and Transwell invasion tests. The expression of related proteins involved in the cell cycle, apoptosis, invasion and signaling pathways was assessed by Western blotting. Results: COX-2 expression was significantly reduced in both breast cancer cell lines infected with Ad-COX-2-shRNA, with obvious inhibition of proliferation, colony forming rate, G2/M phase passage and invasion, as well as induction of apoptosis, in MDA-MB-231 and MCF-7 cells, respectively. At the same time, proteins related to the cell cycle, anti-apoptosis and invasion were significantly downregulated. In addition, c-myc expression and phosphorylation activation of Wnt/${\beta}$-catenin and p38MAPK pathways were reduced by the Ad-COX-2-shRNA. Conclusions: COX-2 expression is associated with proliferation, apoptosis and invasion of breast cancer cells, and its mechanisms of action involve regulating expression of c-myc through the p38MAPK and Wnt/${\beta}$-catenin pathways.

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References

  1. Asara Y, Marchal JA, Carrasco E, et al (2013). Cadmium modifies the cell cycle and apoptotic profiles of human breast cancer cells treated with 5-fluorouracil. Int J Mol Sci, 14, 16600-16. https://doi.org/10.3390/ijms140816600
  2. Bermudez Y, Yang H, Cheng JQ, et al (2008). Pyk2/ERK 1/2 mediate Sp1- and c-myc-dependent induction of telomerase activity by epidermal growth factor. Growth Factors, 26, 1-11. https://doi.org/10.1080/08977190802001389
  3. Bresalier RS, Sandler RS, Quan H, et al (2005). Cardiovascular events associated with rofecoxib in a colorectal adenoma chemoprevention trial. N Engl J Med, 352, 1092-102. https://doi.org/10.1056/NEJMoa050493
  4. Cervello M, Giannitrapani L, La Rosa M, et al (2004). Induction of apoptosis by the proteasome inhibitor MG132 in human HCC cells: Possible correlation with specific caspasedependent cleavage of $\beta$-catenin and inhibition ofb$\beta$-cateninmediated transactivation. Int J Mol Med, 13, 741-8.
  5. Charames G S, Bapat B (2006) Cyclooxygenase-2 knockdown by RNA interference in colon cancer. Int J Oncol, 28, 543-9.
  6. Chen S, Qiong Y, Gardner DG, et al (2006). A role for p38 mitogenactivated protein kinase and c-myc in endothelindependent rat aortic smooth muscle cell proliferation. Hypertension, 47, 252-8. https://doi.org/10.1161/01.HYP.0000198424.93598.6b
  7. Cronin-Fenton DP, Pedersen L, Lash TL, et al (2010). Prescriptions for selective cyclooxygenase-2 inhibitors, non-selective non-steroidal anti-inflammatory drugs, and risk of breast cancer in a population-based case-control study. Breast Cancer Res, 12, 15.
  8. Cusimano A, Fodera D, D'Alessandro N, et al (2007). Potentiation of the antitumor effects of both selective cyclooxygenase-1 and cyclooxygenase-2 inhibitors in human hepatic cancer cells by inhibition of the MEK/ERK pathway. Cancer Biol Therapy, 6, 1461-8.
  9. Daksis JI, Lu RY, Facchini LM, et al (1994). Myc induces cyclin D1 expression in the absence of denovo protein synthesis and links mitogen-stimulated signal transduction to the cell cycle. Oncogene, 9, 3635-45.
  10. Dong JY, Wang Q, Liang QD, et al (2013). Construction and identification of recombinant adenovirus with COX-2- shRNA. Chin J Biologicals, 26, 1072-7.
  11. Fodera D, D'Alessandro N, Cusimano A, et al (2004). Induction of apoptosis and inhibition of cell growth in human hepatocellular carcinoma cells by COX-2 inhibitors. Ann N Y Acad Sci, 1028, 440-9. https://doi.org/10.1196/annals.1322.052
  12. George C (1999). Mechanisms of apoptosis by c-myc. Oncogene, 18, 2967-87. https://doi.org/10.1038/sj.onc.1202727
  13. Half E, Tang XM, Gwyn K, et al (2002). Cyclooxygenase-2 expression in human breast cancers and adjacent ductal carcinoma in situ. Cancer Res, 62, 1676-81.
  14. Han G, Gang W, Yan B, et al (2012). C-myc overexpression promotes osteosarcoma cell invasion via activation of MEKERK pathway. Oncol Res, 20, 149-56. https://doi.org/10.3727/096504012X13522227232237
  15. Harris RE, Beebe-Donk J, Doss H, et al (2005). Aspirin, ibuprofen, and other non-steroidal anti-inflammatory drugs in cancer prevention: a critical review of non-selective COX-2 blockade. Oncol Rep, 13, 559-83.
  16. He TC, Zhou S, Luis T, et al (1998). A simplified system for generating recombinant adenoviruses. Medical Sciences Proc. Natl Acad Sci. 95, 2509-14 https://doi.org/10.1073/pnas.95.5.2509
  17. Hochegger H, Takeda S, Hunt T, et al (2008). Cyclin-dependent kinases and cell cycle transitions: does one fit all? Nat Rev Mol Cell Biol, 9, 910-6.
  18. Hoffman B and Liebermann DA (2008). Apoptotic signaling by c-MYC. Oncogene, 27, 6462-72. https://doi.org/10.1038/onc.2008.312
  19. Janghorban M, Farrell AS, Allen-Petersen BL, et al (2014). Targeting c-MYC by antagonizing PP2A inhibitors in breast cancer. Proc Natl Acad Sci USA, 111, 9157-62. https://doi.org/10.1073/pnas.1317630111
  20. Jovanovic M, Stefanoska I, Radojcic L, et al (2010). Interleukin-8 (CXCL8) stimulates trophoblast cell migration and invasion by increasing levels of matrix metalloproteinase (MMP) 2 and MMP9 and integrins alpha5 and beta1. Reproduction. 139, 789-98. https://doi.org/10.1530/REP-09-0341
  21. Holmes MD, Chen WY, Schnitt SJ, et al (2011). COX-2 expression predicts worse breast cancer prognosis and does not modify the association with aspirin. Breast Cancer Res Treat, 130, 657-62. https://doi.org/10.1007/s10549-011-1651-7
  22. Ferguson RD, Novosyadlyy R, Fierz Y, et al (2012). Hyperinsulinemia enhances c-Myc-mediated mammary tumor development and advances metastatic progression to the lung in a mouse model of type 2 diabetes. Breast Cancer Res, 14, 8.
  23. Komatsu K, Buchanan FG, Katkuri S, et al (2005). Oncogenic potential of MEK1 in rat intestinal epithelial cells is mediated via cyclooxygenase-2. Gastroenterology, 129, 577-90. https://doi.org/10.1016/j.gastro.2005.06.003
  24. Lin MT, Lee RC, Yang PC, et al (2001). Cyclooxygenase-2 inducing Mcl-1-dependent survival mechanism in human lung adenocarcinoma CL1.0 cells. Involvement of phosphatidylinositol 3-kinase/Akt pathway. J Biol Chem, 276, 48997-9002. https://doi.org/10.1074/jbc.M107829200
  25. Llovet JM, Bruix J (2008). Molecular targeted therapies in hepatocellular carcinoma. Hepatol, 48, 1312-27. https://doi.org/10.1002/hep.22506
  26. Murata H, Kawano S, Tsuji S, et al (1999). Cyclooxygenase-2 overexpression enhances lymphatic invasion and metastasis in human gastric carcinoma. Amer J Gastroenterol, 94, 451-5. https://doi.org/10.1111/j.1572-0241.1999.876_e.x
  27. Parkin DM, Bray F, Ferlay J, et al (2005). Global cancer statistics, 2002. CA Cancer J Clin, 55, 74-108 https://doi.org/10.3322/canjclin.55.2.74
  28. Patel VA, Dunn MJ, Sorokin, et al (2002). Regulation of MDR-1 (P-glycoprotein) by cyclooxygenase-2. J Biol Chem, 277, 38915-20. https://doi.org/10.1074/jbc.M206855200
  29. Schutze N (2004). siRNA technology. Mol Cell Endocrinol, 213, 115-9. https://doi.org/10.1016/j.mce.2003.10.078
  30. Shalaby MA, Nounou HA, Alanazi MS, et al (2014). Associations between single nucleotide polymorphisms of COX-2 and MMP-2 genes and colorectal cancer susceptibility in the Saudi population. Asian Pac J Cancer Prev, 15, 4989-94. https://doi.org/10.7314/APJCP.2014.15.12.4989
  31. Shirahama T, Arima J, Akiba S, et al (2001). Relation between cyclooxygenase-2 expression and tumor invasiveness and patient survival in transitional cell carcinoma of the urinary bladder. Cancer, 92, 188-93. https://doi.org/10.1002/1097-0142(20010701)92:1<188::AID-CNCR1308>3.0.CO;2-W
  32. Siegfried JM, Gubish CT, Rothstein ME, et al (2007). Signaling pathways involved in cyclooxygenase-2 induction by hepatocyte growth factor in non small-cell lung cancer. Mol Pharmacol, 72, 769-79. https://doi.org/10.1124/mol.107.034215
  33. Singh B, Berry JA, Shoher A, et al (2006). COX-2 overexpression increases motility and invasion of breast cancer cells. Int J Oncol, 26, 1393-9.
  34. Singh B, Cook KR, Vincent L, et al (2011). Role of COX-2 in tumorospheres derived from a breast cancer cell line. J Surg Res, 168, 39-49. https://doi.org/10.1016/j.jss.2010.03.003
  35. Sobolewski C, Cerella C, Dicato M, et al (2010). The role of cyclooxygenase-2 in cell proliferation and cell death in human malignancies. Int J Cell Biol, 2010, 215158 (21 pages).
  36. Song D, Miao Cui M, Zhao G, et al (2014). Pathway-based analysis of breast cancer. Am J Transl Res, 6, 302-11.
  37. Strillaeeit A, Griffoni C, Spiral E, et al (2006). RNA interference as a key to knockdown overexpressed cyclooxygenase-2 in tumor cells. Br J of Cancer, 94, 1300-10. https://doi.org/10.1038/sj.bjc.6603094
  38. Su B, Chen S (2009). Lead optimization of COX-2 inhibitor nimesulide analogs to overcome aromatase inhibitor resistance in breast cancer cells. Bioorg Med Chem Lett, 19, 6733-5. https://doi.org/10.1016/j.bmcl.2009.09.109
  39. Tai WT, Cheng AL, Shiau CW, et al (2011). Signal transducer and activator of transcription 3 is a major kinase-independent target of sorafenib in hepatocellular carcinoma. J Hepatol, 55, 1041-8. https://doi.org/10.1016/j.jhep.2011.01.047
  40. Xia JJ, Pei LB, Zhuang JP, et al (2010). Celecoxib inhibits beta-catenin-dependent survival of the human osteosarcoma MG-63 cell line. J Int Med Res, 38, 1294-304. https://doi.org/10.1177/147323001003800411
  41. Yamanaka Y, Shiraki K, Inoue T, et al (2006). COX-2 inhibitors sensitize human hepatocellular carcinoma cells to TRAILinduced apoptosis. Int J Mol Med, 18, 41-7.

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