Advertisement

Chinese Journal of Cancer Research

, Volume 15, Issue 2, pp 86–91 | Cite as

Investigation of the therapeutic effect of expression of trailin vivo on mouse hepatocellular carcinoma

  • Zhang Gui-mei 
  • Xue Sheng-li 
  • Zhang Hui 
  • Li Pei-yuan 
  • Li Dong 
  • Feng Zuo-hua 
Article
  • 17 Downloads

Abstract

Objective: To construct an eukaryotic expressing plasmid of mouse TRAIL (mTRAIL), and investigate its ability to induce the apoptosis of hepatocellular carcinoma cellsin vitro and in vivo, its inhibitory effect on the growth of hepatocellular carcinoma, and its synergism with pCH510, an eukaryotic expressing plasmid of recombinant human FN polypeptide. Methods: The eukaryotic expressing plasmid of mTRAIL was constructed by RT-PCR and DNA recombination techniques. Gene transfection was performedin vitro and in vivo. The apoptosis rate of hepatocellular carcinoma cells was measured by Flow Cytometry. The apoptosis of hepatocellular carcinoma cells was detected by TdT-mediated dUTP nick end labeling (TUNEL) and histochemistry techniques. The inhibitory effect of gene transfection on solid tumor was observed in mice. Results: The cDNA of mTRAIL was amplified by RT-PCR from the RNA of mouse spleen cells, and cloned into the eukaryotic expressing vector pcDNA3.1. The recombinant plasmid was designated as pXl. The BHK cells transfected with plasmid pXl could attack H22 hepatocellular carcinoma cells and induce the apoptosis of them. The transfection of plasmid pX1 through injection into mouse muscles could inhibit the growth of hepatocellular carcinoma by inducing the apoptosis of tumor cells. Plasmid pX1 and pCH510 had a synergistic inhibitory effect on the hepatocellular carcinoma growth. Conclusion: Plamid pX1 could be expressed in cells andin vivo in mouse. The expression of pX1in vivo andin vitro could induce the apoptosis of hepatocellular carcinoma cells and inhibit the growth of hepatocellular carcinoma. Plasmid pX1 and pCH510 had a synergistic inhibitory effect on the hepatocellular carcinoma growth.

Key words

TRAIL Eukaryotic Expression Tumor therapy 

CLC number

R73-36+2 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. [1]
    Wiley SR, Schooley K, Smolak PJ,et al. Identification and characterization of a new member of the TNF family that induces Apoptosis[J]. Immunity 1995; 3: 673–82.PubMedCrossRefGoogle Scholar
  2. [2]
    Pitti RM, Marsters SA, Ruppert S,et al. Induction of Apoptosis by Apo-2 ligand, a new member of the tumor necrosis factor cytokine family[J]. J Biol Chem 1996; 271: 12687–90.PubMedCrossRefGoogle Scholar
  3. [3]
    Gura T. How TRAIL kills cancer cells, but not normal cells[J]. Science 1997; 277: 768.PubMedCrossRefGoogle Scholar
  4. [4]
    Ye Shiqiao, Feng Zuohua, Li Dong, et al. Construction and expression of eukaryotic expressing vector pCH510 of polypeptide CH50 and its chemotaxis and antitumor function by transfectionin vivo(in Chinese)[J]. Chin J Cancer Biother 2001; 8: 23–6.Google Scholar
  5. [5]
    Sambrook J, Fritsch EF, Maniatis T. Molecular Cloning: A Laboratory Manual[M]. 2nd ed. New York: Cold Spring Harbor Laboratory Press, 1989. P???Google Scholar
  6. [6]
    Vassalli P. The pathophysiology of tumor necrosis factors[J] Annu. Rev Immunol. 1992; 10: 411–52.PubMedCrossRefGoogle Scholar
  7. [7]
    Ogasawara J, Watanabe-Fukunaga R, Adachi M, et al. Lethal effect of the anti-Fas antibody in mice[J]. Nature 1993; 364: 806–9.PubMedCrossRefGoogle Scholar
  8. [8]
    Walczak H, Miller RE, Ariail K,et al. Tumoricidal activity of TNF-related Apoptosis inducing ligandin vivot[J]. Nature Med 1999; 5: 157–63.PubMedCrossRefGoogle Scholar
  9. [9]
    Pan G, O’Rourke K, Chinnaiyan AM, et al. The receptor for the cytotoxic ligand TRAIL[J]. Science 1997; 276: 111–3.PubMedCrossRefGoogle Scholar
  10. [10]
    Pan G, Ni J, Wei Y, et al. An antagonist decoy receptor and a death domain-containing receptor for TRAIL[J]. Science 1997; 277: 815–7.PubMedCrossRefGoogle Scholar
  11. [11]
    Sheridan JP, Marsters SA, Pitti RM, et al. Control of TRAIL-induced Apoptosis by a family of signaling and decoy receptors [J]. Science 1997; 277: 818–21.PubMedCrossRefGoogle Scholar
  12. [12]
    Emery JG, McDonnell P, Burke MB, et al. Osteoprotegerin is a receptor for the cytotoxic ligand TRAIL[J]. J Biol Chem 1998; 273: 14363–7.PubMedCrossRefGoogle Scholar
  13. [13]
    Schulze-Osthoff K, Ferrari D, Los M, et al. Apoptosis signaling by death receptors [J]. Eur J Biochem 1998; 254:439–59.PubMedCrossRefGoogle Scholar
  14. [14]
    Rosenberg SA. Progress in human tumor immunology and immunotherapy[J]. Nature, 2001, 411: 380–384.PubMedCrossRefGoogle Scholar
  15. [15]
    Sun Hong, Zhang Xiyin, Feng Youji. Investigation on the therapeutic effect of recombinant interleukin 2 in combination with chemotherapy on relapse and metastasis of oophoroma(in Chinese)[J]. Chin J Cancer Biother 1999; 6: 312–3.Google Scholar
  16. [16]
    Huang Bo, Feng Zuohua, Zhang Guimei, et al. The therapeutic effect of eukaryotic expressing vector pCH510 of recombinant polypeptide of FN linking chemotherapy on tumor-bearing mice(in Chinese)[J]. Chin J Cancer Biother, 2001, 8: 168–172.Google Scholar
  17. [17]
    Doherty DE, Henson PM, Clark RA. Fibronectin fragment containing the RGDS cell-binding domain mediate monocyte migration into the rabbit lung. A potential mechanism for C5 fragment-induced monocyte lung accumulation[J]. J Clin Invest 1990; 86: 1065–75.PubMedCrossRefGoogle Scholar

Copyright information

© Chinese Journal of Cancer Research 2003

Authors and Affiliations

  • Zhang Gui-mei 
    • 1
  • Xue Sheng-li 
    • 1
  • Zhang Hui 
    • 1
  • Li Pei-yuan 
    • 1
  • Li Dong 
    • 1
  • Feng Zuo-hua 
    • 1
  1. 1.Department of Biochemistry and Molecular Biology, Tongji Medical CollegeHuazhong University of Science and TechnologyWuhan

Personalised recommendations