Gene Transfer Mediated Through Adeno-associated Virus InducesKiSS-1 In Adult MG-63 Cells

[Objective]The aim of this study is to construct vectors based on adeno-associated virus type2 (rAAV2) carrying KiSS-1 and observe their expression in MG63 cell lines.To investigate the proliferation,adhesion and metastasis ability of MG63 cells in vitro after gene transfection for gene therapy of osteosarcoma.[Methods]1. To construct vectors based on adeno-associated virus carrying KiSS-1.2. To transfect KiSS-1 into MG63 cell lines through lipofectamine2000.3. To detecte the expression of KiSS-1mRNA and metastin in the transfected cell lines by RT-PCR and western blot.4. To examine the effect of the transfection in vitro by cell cell proliferation.5. To test the difference of adhesion,invasion and metastasis ability between the transfected and non-transfected cell lines by use of transwell-room,millicell-room and tumor adhesion experiment.[Results]1. KiSS-1 was extract from the placenta and connected into the vector PSNAV2.0 through T vector. The result was correct by sequencing.2. The expression of KiSS-1 between transfected and non-transfected cell lines was observed and considered to be a significant difference.3. The cell proliferation was observed to be slow down.4. The adhesion ability was inhibit in the transfected cell lines.5. The invasion ability of transfected cell lines was lower than the non-transfected cell lines.6. The metastasis ability of transfected cell lines was lower than the non-transfected cell lines.[Conclusions]1. The way that transfer KiSS-1 into the MG63 cell lines by lipofectamine2000 is safe,high-efficiency and low toxicity.2. The PSNAV2.0-KiSS-1 plasmid can express the target gene in high Level.3. The cell proliferation was observed to be slow down.4. The adhesion,invasion and metastasis ability of transfectd cell lines decresed significantly.5. KiSS-1 is a promising candidate tumor-suppressor gene and may play a key role in the metastasis.