Effect Of KiSS-1 Gene On The Invasive Ability Of Differently Malignant Osteosarcoma Cell In Vitro

[Objective]The aim of this study is to explore the effect of KiSS-1 gene on differently malignant osteoarcoma cells in vitro and to investigate the proliferation,migration and invasive ability of MG-63 and U-2OS cells in vitro after KiSS-1 gene transfection.The expression of matrix metallproteinases and tissue inhibitors of metallprotenises were examined to explore the molecular mechanism of KiSS-1 gene antimetastasis capability.[Methods]1 To transfect KiSS-1 gene into MG-63 and U2-OS cells through lipofectamine2000.2. To confirm the expression of KiSS-1gene at the level of the transcription and translation in the transfected cell lines by RT-PCR and western blot respectively.3. To investigate the difference of proliferation,migration and invasive ability between the transfected and non-transfected cell lines by CCK-8,Wound healing assay andTranswell.4. The expression of MMP and TIMP was evaluated by sybr-green realtime PCR before and after gene transfection by real time PCR.5. To investigate the amount of the Pro-MMP and the active MMP by gelatin zymography before and after transfection.6. Detect the change of TIMPs between the transfected and non-transfected cell lines by Western blot .[Results]1 The MG63 and U2-OS cell line with stable expression of KiSS-1 were selected by G418 transfected with KiSS-1 by lipofectamine. 2.The expression of KiSS-1 between transfected and non-transfected celllines was observed and considered to be a significant difference.3 The cell proliferation migration and invisive ability of transfected celllines was observed to be slow down .4 The expression of MMP-2 and active MMP-2 decreased aftertransfetion in MG-63 cell; the expression of MMP-9 decrease after thetransfection.5 The expression of TIMP1 up-regulated significantly after transfection inboth MG63 and U2OS cell., but no significance observed in theexpression of TIMP2.[Conclusions]1 The way that transfer KiSS-1 into the MG63,U-2OS cell lines bylipofectamine2000 is safe,high-efficiency and low toxicity2 The PSNAV2.0-KiSS-1 plasmid can express the target gene in high level.3 The cell proliferation migration and invasive ability of transfectd cell linesdramatically attenuate.4 The expression of the MMP2,MMP9 and TIMP1 may be affected by the Theexpression of Kiss-1 ,thus the invasion was reducued5 The Kiss-1 may be involve in the process of invasion and metastasisinosteosarcoma, and would be a promising candidate tumor-suppressor gene andplay a key role in the metastasis in the future.