The Prelimimary Study Of The Expression And Function Of VEGFR-1 In Hepatocellular Carcinoma Cells

【Objective】The role of vascular endothelial growth factor(VEGF) and its receptors participating in the pathological blood vessel formation has been widely known.In recent years,with the deep research,people has paid more attention to the non-angiogenesis-promoting fuction of VEGFR-1 on tumor invasion and progession.The in vivo or in vitro study of colon cancer,pancreatic carcinoma,breast cancer has shown that tumor tissues and cell lines both express functional VEGFR-1,and the activation of VEGFR-1 can inhance tumor invasion and metastasis by inducing the epithelial cells change to the stroma cells(EMT) and promoting the secretion and active function of MMPs. Hepatocellular carcinoma(HCC) is the solid tumor rich in the expression of VEGF,blood vessel and invasion.Up to now,there is no systematic study about the expression,function and mechanism of VEGFR-1 in HCC at home and abroad.In this study,we detected the expression of VEGFR-1 protein in different hepatocarcinoma cell lines,we also detected the expression of nuclear transcription factor:Snail protein and the functional changes of MMP-9 in cell culture supernatant which were both stimulated by VEGF-B167 from SMMC-7721 cell line;at the same time we detected the expression of Snail protein in different invasive cell lines:MHCC-97H,SMMC-7721,HepG2 and the normal liver cell line:LO2. To make the preliminary research of the function and mechanism of VEGFR-1 in hepatocellular carcinoma cells,and to provide a new theoretical basis on the prevention and treatment of hepatocellular carcinoma in future.[Methods]Culture various cell lines:LO2,HepG2,SMMC-7721,MHCC-97H.To detect VEGFR-1,Snai protein expression by Western Blot (WB),.SMMC-7721 cell line were renewed by non-FBS 1640 medium,starvation overnight,the next day by adding VEGF-B167,at the same time set the IgG isotype control and blank control, to detect the Snail protein expression of different treatment groups and Snail protein expression changes stimulated by VEGF-B167 at different time points(0h,8h,24h,28h,32h) by WB; to detect the functional activity of MMP-9 in SMMC-7721 cell line supernant treated by VEGF-B167,IgG isotype control and blank control through gelatin zymography. At last for statistical analysis.[Results]1,VEGFR-1 protein expressed in LO2,HepG2,SMMC-7721,MHCC-97H cell lines,semiquantitative gray scale analysis show that the highest expression of VEGFR-1 in MHCC-97H cell,the moderate in SMMC-7721, while the lowest in LO2. 2,MHCC-97H,SMMC-7721,HepG2 hepatoma cell lines expressed Snail protein and LO2 did not expression Snail. semiquantitative gray scale analysis show that the highest expression of Snail in MHCC-97H cell,the moderate in SMCC-7721, while the lowest in HepG2.3,At 28h the protein expression of Snail is to the peak by the stimulation of VEGF-B167 to VEGFR-1 in SMMC-7721cell line,while at 32h the snail expression gradually dreased. VEGF-B167 stimulation group: IgG isotype contol4, The expression of Snail was significantly increased by the stimulation of VEGF-B167 to VEGFR-1 in SMMC-7721 cell line at 28h as to IgG isotype control,the blank group. 5,The active function of MMP-9 was significantly increased by the stimulation of VEGF-B167 to VEGFR-1 in SMMC-7721 cell line at 28h as to IgG isotype control,the blank group by gelatin-zymogram.The MMP-2 band (64KD) is unnoticeable.6,The protein expression of MMP-9 in SMMC-7721 cell line at the different time points(0h,24h,28h,32h,48h).24h the band appeared,with time,the expression of MMP-9 gradually inceased,48h the band disappeared.[Conclusions]VEGFR-l expressed in hepatoma cell lines and its activation can inhance the invasive activity of cell lines through the increased expression and enhanced function of MMP-9,while Snail participated and mediated in this process.