DOI QR코드

DOI QR Code

Combined EGFR and c-Src Antisense Oligodeoxynucleotides Encapsulated with PAMAM Denderimers Inhibit HT-29 Colon Cancer Cell Proliferation

  • Nourazarian, Ali Reza ;
  • Najar, Ahmad Gholamhoseinian ;
  • Farajnia, Safar ;
  • Khosroushahi, Ahmad Yari ;
  • Pashaei-Asl, Roghiyeh ;
  • Omidi, Yadollah
  • Published : 2012.09.30

Abstract

Colon cancer continues to be one of the most common cancers, and the importance and necessity of new therapies needs to be stressed. The most important proto-oncogen factors for colon cancer appear to be epidermal growth factor receptor, EGFR, and c-Src with high expression and activity leading to tumor growth and ultimately to colon cancer progression. Application of c-Src and EGFR antisense agents simultaneously should theoretically therefore have major benefit. In the present study, anti-EGFR and c-Src specific antisense oligodeoxynucleotides were combined in a formulation using PAMAM dendrimers as a carrier. Nano drug entry into cells was confirmed by flow cytometry and fluorescence microscopy imaging and real time PCR showed gene expression of c-Src and EGFR, as well as downstream STAT5 and MAPK-1 with the tumor suppressor gene P53 to all be downregulated. EGFR and c-Src protein expression was also reduced when assessed by western blotting techniques. The effect of the antisense oligonucleotide on HT29 cell proliferation was determined by MTT assay, reduction beijng observed after 48 hours. In summary, nano-drug, anti-EGFR and c-Src specific antisense oligodeoxynucleotides were effectively transferred into HT-29 cells and inhibited gene expression in target cells. Based on the results of this study it appears that the use of antisense EGFR and c-Src simultaneously might have a significant effect on colon cancer growth by down regulation of EGFR and its downstream genes.

Keywords

Colon cancer;HT-29;antisense;EGFR;c-Src;PAMAM dendrimers

References

  1. Albertazzi L,SerresiI M, Albanese A, Beltram F (2010). Dendrimer internalization and intracellylar trafficking in living cells. Mol Pharm, 7, 680-8. https://doi.org/10.1021/mp9002464
  2. Beale G, HolliIns AJ, Benboubetra M, et al (2003). Gene silencing nucleic acids designed by scanning arrays:Anti- EGFR activity of siRNA, ribozyme and DNA enzymes targeting a single Hybridization-accessible region using the same delivery system. J Drug Targeting, 7, 449-56.
  3. Bowles TL, Parsons C, Muilenburg D, Bold RJ (2009). Targeted inhibition of AKT in pancreatic cancer. Curr Cancer Therapy Rev, 5, 288-95. https://doi.org/10.2174/157339409789712654
  4. Bradford M (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
  5. Chapnick D, Warner L, Bernet J, RAO T, Liu X (2011). The TGFbeta and MAPK pathways in cancer progression. Cell and Biosci, 1, 42. https://doi.org/10.1186/2045-3701-1-42
  6. Eder I, Haag P, Bartsch G, Klocker H (2005). Gene therapy strategies in prostate cancer. Curr Gene Ther, 5, 1-10. https://doi.org/10.2174/1566523052997424
  7. Ellis LM, Staley CA, LIU W, et al (1998). Down-regulation of vascular endothelial growth factor in a human colon carcinoma cell line transfected with an antisense expression vector specific for c-src. J Biol Chem, 273, 1052-7. https://doi.org/10.1074/jbc.273.2.1052
  8. FINN R (2008). Targeting Src in breast cancer. Annals Oncol, 19, 1379-86. https://doi.org/10.1093/annonc/mdn291
  9. Irby RB, Yeatman T (2000). Role of Src expression and activation in human cancer. Oncogene, 19, 5636-42. https://doi.org/10.1038/sj.onc.1203912
  10. Kopetz S (2007). Targeting SRC and epidermal growth factor receptor in colorectal cancer: rationale and progress into the clinic. Gastrointest Cancer Res, 1, 37-41.
  11. Kumar R, SriniIvasan S, Pahari P, Rohr J, Damodaran C (2010). Activating stress-activated protein kinase-mediated cell death and inhibiting epidermal growth factor receptor signaling: a promising therapeutic strategy for prostate cancer. Mol Cancer Ther, 9, 2488-96. https://doi.org/10.1158/1535-7163.MCT-10-0180
  12. LIEU C, Kopetz S (2010). The SRC family of protein tyrosine kinases: a new and promising target for colorectal cancer therapy. Clin Colorectal Cancer, 9, 89-94. https://doi.org/10.3816/CCC.2010.n.012
  13. Mukhopadhyay D, Tsiokas L, Zhou XM , et al (1995). Hypoxic induction of human vascular endothelial growth factor expression through c-Src activation. Nature, 375, 577-81. https://doi.org/10.1038/375577a0
  14. Nakhlband A, Barar J, Bidmeshkipour A, Heidari H, Omidi Y (2010). Bioimpacts of anti epidermal growth factor receptor antisense complexed with polyamidoamine dendrimers in human lung epithelial adenocarcinoma cells. J Biomed Nanotechnol, 6 360-9. https://doi.org/10.1166/jbn.2010.1131
  15. Nourazarian AR, Pashaie AR, Omidi Y, Gholamhoseinian NA(2012). c-Src antisense complexed with PAMAM denderimes decreases of c-Src expression and EGFRdependent downstream genes in the human HT-29 colon cancer cell line. Asian Pac J Cancer Prev, 13, 2235-40. https://doi.org/10.7314/APJCP.2012.13.5.2235
  16. Omidi Y, Barar J (2009). Induction of human alveolar epithelial cell growth Factor receptors by dendrimeric nanostructure. Int J Toxicol, 28, 113-22. https://doi.org/10.1177/1091581809335177
  17. Sabbah M, Emami S, Redeuilh G, et al (2008). Molecular signature and therapeutic perspective of the epithelialto- mesenchymal transitions in epithelial cancers. Drug Resistance Updates, 11, 123-51. https://doi.org/10.1016/j.drup.2008.07.001
  18. Summy JM, Gallick GE (2003). Src family kinases in tumor progression and metastasis. Cancer Metastasis Rev, 22, 337-58. https://doi.org/10.1023/A:1023772912750
  19. Summy JM, Trevino JG, Lesslie DP, et al (2005). AP23846, a novel and highly potent Src family kinase inhibitor, reduces vascular endothelial growth factor and interleukin-8 expression in human solid tumor cell lines and abrogates downstream angiogenic processes. Mol Cancer Ther, 4, 1900-11. https://doi.org/10.1158/1535-7163.MCT-05-0171
  20. Talamonti M, Roh M, Curley S, Gallick G (1993). Increase in activity and level of pp60c-src in progressive stages of human colorectal cancer. J Clin Invest, 91, 53-60. https://doi.org/10.1172/JCI116200
  21. Xiong H, Su WY, Liang QC, et al (2009). Inhibition of STAT5 induces G1 cell cycle arrest and reduces tumor cell invasion in human colorectal cancer cells. Lab Invest, 89, 717-25. https://doi.org/10.1038/labinvest.2009.11
  22. Yamaguchi M, Tanaka T, Waki M, et al (1997). Antisense src expression inhibits tyrosine phosphorylation of Shc and its association with Grb2 and Sos which leads to MAP kinase activation in U937 human leukemia cells. Leukemia, 11, 497-503. https://doi.org/10.1038/sj.leu.2400605
  23. Yavari K, Taghikhani M, Maragheh MG, Mesbahnamin SA, BabaieI MH (2009). Knockdown of IGF-IR by RNAi inhibits SW480 colon cancer cells growth in vitro. Arch Med Res, 40, 235-40. https://doi.org/10.1016/j.arcmed.2009.03.001

Cited by

  1. Thymoquinone induces apoptosis in human colon cancer HCT116 cells through inactivation of STAT3 by blocking JAK2- and Src-mediated phosphorylation of EGF receptor tyrosine kinase vol.32, pp.2, 2014, https://doi.org/10.3892/or.2014.3223
  2. Strategy for selecting nanotechnology carriers to overcome immunological and hematological toxicities challenging clinical translation of nucleic acid-based therapeutics vol.12, pp.7, 2015, https://doi.org/10.1517/17425247.2015.1042857
  3. Combinatorial nanomedicines for colon cancer therapy vol.8, pp.1, 2015, https://doi.org/10.1002/wnan.1353
  4. Potential Molecular Targets in the Treatment of Lung Cancer Using siRNA Technology vol.36, pp.1, 2018, https://doi.org/10.1080/07357907.2017.1416393
  5. Natural low- and high-density lipoproteins as mighty bio-nanocarriers for anticancer drug delivery vol.82, pp.3, 2018, https://doi.org/10.1007/s00280-018-3626-4