Fused Polypeptide with DEF Induces Apoptosis of Lung Adenocarcinoma Cells

  • Liang, Ai-Ling (Key Laboratory for Clinical Laboratory Diagnostics of Education Ministry of China and School of Clinical Diagnostic and Laboratory Medicine, Chongqing Medical University) ;
  • Zhang, Ting-Ting (Medical Molecular Diagnostics Key Laboratory of Guangdong) ;
  • Zhou, Ning (Medical Molecular Diagnostics Key Laboratory of Guangdong) ;
  • Huang, Di-Nan (Medical Molecular Diagnostics Key Laboratory of Guangdong) ;
  • Liu, Xin-Guang (Medical Molecular Diagnostics Key Laboratory of Guangdong) ;
  • Liu, Yong-Jun (Medical Molecular Diagnostics Key Laboratory of Guangdong) ;
  • Tu, Zhi-Guang (Key Laboratory for Clinical Laboratory Diagnostics of Education Ministry of China and School of Clinical Diagnostic and Laboratory Medicine, Chongqing Medical University)
  • Published : 2013.12.31


To analyze the effects of a new unknown peptide DEF on the growth of tumor cells, a fused polypeptide TAT-DV1-DEF was designed and synthesized. The lung adenocarcinoma cell line GLC-82 treated with TAT-DV1-DEF was analyzed with a cell counting kit 8, and the location of polypeptides in cells was observed under laser confocal microscopy. The efficiency of polypeptide transfection and changes in nuclear morphology were analyzed by flow cytometry and fluorescence microscopy, respectively. Finally, the mechanism of tumor cell growth inhibition was evaluated by Western blotting. We found that TAT-DV1-DEF could significantly inhibit the growth of the lung adenocarcinoma cell line GLC-82, but not the normal human embryonic kidney cell line HEK-293. Polypeptides were found to be mostly localized in the cytoplasm and some mitochondria. The efficiency of polypeptide transfection in the two cell types was approximately 99%. Apoptotic nuclei were observed under fluorescence microscopy upon treatment with polypeptides and DAPI staining. Western blot analyses indicated that the polypeptide inhibition of tumor cell growth was apoptosis dependent. In the present study, we demonstrated that fused polypeptides could induce apoptosis of the lung adenocarcinoma cell line GLC-82, indicating that the new unknown peptide DEF has antitumor effects.


  1. Anderson WF, Blaese RM, Culver K (1990). The ada human gene therapy clinical protocol: Points to consider response with clinical protocol. Hum Gene Ther, 1, 331-62.
  2. Balkwill F (2004). The significance of cancer cell expression of the chemokine receptor cxcr4. Semin Cancer Biol, 14, 171-9.
  3. Ceron JM, Contreras-Moreno J, Puertollano E, et al (2010). The antimicrobial peptide cecropin a induces caspaseindependent cell death in human promyelocytic leukemia cells. Peptides, 31, 1494-503.
  4. Hao H, Dong Y, Bowling MT, et al (2007). E2F-1 induces melanoma cell apoptosis via puma up-regulation and bax translocation. BMC Cancer, 7, 24.
  5. Hsu JC, Lin LC, Tzen JT, et al (2011). Characteristics of the antitumor activities in tumor cells and modulation of the inflammatory response in raw264.7 cells of a novel antimicrobial peptide, chrysophsin-1, from the red sea bream (chrysophrys major). Peptides, 32, 900-10.
  6. Huerta S, Goulet EJ, Livingston EH (2006). Colon cancer and apoptosis. Am J Surg, 191, 517-26.
  7. Jadus MR, Natividad J, Mai A, et al (2012). Lung cancer: A classic example of tumor escape and progression while providing opportunities for immunological intervention. Clin Dev Immunol, 2012, 160724-44.
  8. Kashiwagi H, Mcdunn JE, Goedegebuure PS, et al (2007). Tatbim induces extensive apoptosis in cancer cells. Ann Surg Oncol, 14, 1763-71.
  9. Liang A, Liu Y, Hou G, et al (2012). Study of screening for defensing genes in scallop by improved rapid amplification of cdna ends. Lab Med Clin, 9, 1160-2.
  10. Liang MD (1985). Establishment of lung adenocarcinoma cell line in gejiu (glc-82) and study of its biologic properties. Zhonghua Zhong Liu Za Zhi, 7, 81-2.
  11. Lin HJ, Huang TC, Muthusamy S, et al (2012). Piscidin-1, an antimicrobial peptide from fish (hybrid striped bass morone saxatilis x m. Chrysops), induces apoptotic and necrotic activity in ht1080 cells. Zoolog Sci, 29, 327-32.
  12. Liu Y, Li Y, Wang H, et al (2009). Bh3-based fusion artificial peptide induces apoptosis and targets human colon cancer. Mol Ther, 17, 1509-16.
  13. Marshall E (2011). Cancer research and the $90 billion metaphor. Science, 331, 1540-1.
  14. Martin E, Ganz T, Lehrer RI (1995). Defensins and other endogenous peptide antibiotics of vertebrates. J Leukoc Biol, 58, 128-36.
  15. Movassaghian S, Moghimi HR, Shirazi FH, et al (2013). Efficient down-regulation of pkc-alpha gene expression in a549 lung cancer cells mediated by antisense oligodeoxynucleotides in dendrosomes. Int J Pharm, 441, 82-91.
  16. Nagahara H, Vocero-Akbani AM, Snyder EL, et al (1998). Transduction of full-length tat fusion proteins into mammalian cells: Tat-p27kip1 induces cell migration. Nat Med, 4, 1449-52.
  17. Snyder EL, Meade BR, Saenz CC, et al (2004). Treatment of terminal peritoneal carcinomatosis by a transducible p53-activating peptide. PLoS Biol, 2, E36.
  18. Snyder EL, Saenz CC, Denicourt C, et al (2005). Enhanced targeting and killing of tumor cells expressing the cxc chemokine receptor 4 by transducible anticancer peptides. Cancer Res, 65, 10646-50.
  19. Tian Y, Wang H, Li B, et al (2013). The cathelicidin-bf lys16 mutant cbf-k16 selectively inhibits non-small cell lung cancer proliferation in vitro. Oncol Rep, 30, 2502-10.
  20. Van ZH, Carpentier G, Dos SC, et al (2012). Antitumor and angiostatic activities of the antimicrobial peptide dermaseptin b2. PLoS One, 7, e44351.
  21. Wu SP, Huang TC, Lin CC, et al (2012). Pardaxin, a fish antimicrobial peptide, exhibits antitumor activity toward murine fibrosarcoma in vitro and in vivo. Mar Drugs, 10, 1852-72.
  22. Yu J, Yue W, Wu B, et al (2006). Puma sensitizes lung cancer cells to chemotherapeutic agents and irradiation. Clin Cancer Res, 12, 2928-36.