JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES) ›› 2012, Vol. 50 ›› Issue (7): 41-.

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Construction of a recombinant adenoassociated viral vector expressing SAC and its inhibition on CAM xenograft prostate cancer

YU Zhang-jian1, ZHANG Shi-bao1, LIU Qing-yong1, RUAN Xi-yun2, YANG Guang-xiao3, WANG Quan-ying3   

  1. 1. Department of Urology, Jinan Central Hospital Affiliated to Shandong University, Jinan 250013, China;
    2. Department of Neurology, Provincial Hospital Affiliated to Shandong University, Jinan 250021, China;
    3. Xi’an Huaguang Biological Engineering Limited Company, Xi’an 710025, China
  • Received:2011-12-02 Online:2012-07-10 Published:2012-07-10

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Abstract:

 Objective   To construct a recombinant adeno-associated virus expression plasmid containing a unique core domain of par-4 sufficient for selective apoptosis induction in cancer cells (SAC) and protein transduction domain TAT, and investigate the effect of the SAC on chick chorioallantoic membrane (CAM) xenograft prostate cancer. Methods   Plasmids pAAV/Ad, pFG140 and pSSCMV/NT4-SAC-HA2-TAT were transfected into 293 cells by method of Ca3(P04)2 to harvest rAAV-NT4-SAC-HA2-TAT whose titer was detected by dot blot. PC-3 cells were grafted on CAM in the 9th day of the embryonic development to construct the CAM xenograft tumor model of prostate cancer. The growth of xenograft tumor and richness of CAM capillaries around tumor were observed and photographed. 3 days later， those chick embryos with tumor formation were randomly divided into three groups (the PBS group, the AAV group，and the rAAV group). The tumors of different groups were compared on the volume 4 days after the treatment and fixed with formalin for further analyzation using standard histopathology. The expression of SAC was detected with immunofluorescence method. Results   Recombinant plasmid pSSCMV/NT4-SAC-HA2-TAT was successfully constructed confirmed by restriction enzyme digestion. The viral titer was about 3.34×1010～3.34×1011cfu/mL detected by dot blot. The expression of SAC fusion peptide in the tumor tissue was detected by immunefluorescence test. The xenograft tumor could grow on the CAM, and a mass of capillaries around the tumor were observed. Tumors in the rAAV group significantly shrank. The invasiveness of PC-3 cells was inhibited and the tumor cells distributions were limited under the microscope.  Conclusion   The xenograft tumor model of prostate cancer on CAM is established successfully. Fusion peptide of SAC plays a significant role in inhibiting the invasiveness and inducting the apoptosis of PC-3 cells.

Key words: Par-4-SAC; Recombinant adeno-associated virus; Chick chorioallantoic membrane; Androgenindependent prostate cancer; Xenograft tumor; Gene therapy

CLC Number: 

  • R737.25
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