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《Chinese Journal of Pathophysiology》 2011-07
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Effect of CD97 gene silencing by small interfering RNA on migration and invasion of gastric carcinoma cell lines

SHI Xiao-yu1,TAO Si-feng2,CHEN Li1,PENG Shu-you1,HOANG-VU Cuong3 (1Department of General Surgery,2Department of Oncosurgery,The Second Affiliated Hospital,Zhejiang University School of Medicine,Hangzhou 310009,China.3Department of Surgery,Martin Luther University of Halle-Wittenberg,Halle 06099,Germany.)  
AIM: To investigate the effect of CD97 gene silencing by small interfering RNA(siRNA) on migration and invasion of gastric carcinoma cell lines.METHODS: Gastric carcinoma cell lines AGS and MGC803 were used in the study.Four pairs of siRNA were designed according to the sequence of CD97 gene and synthesized chemically.The siRNAs were transfected into the gastric carcinoma cell lines.Forty-eight hours after transfection,the total RNA was extracted and the mRNA expression of CD97 was detected by real-time RT-PCR so as to screen the most effective siRNA.The protein level of CD97 was also measured by fluorescence-activated cell sorting(FACS) 72 h after Transfection.The abilities of migration and invasion were evaluated by Transwell test.The viability of the cells was measured by MTT method.RESULTS: Real-time RT-PCR and FACS revealed that CD97-siRNA notably down-regulated CD97 expression at both mRNA and protein levels.The mRNA level decreased by(89.34±9.95)% and(95.42±1.93)% in AGS and MGC803 cells,respectively.The protein levels of CD97EGF and CD97stalk in AGS cells decreased by(19.29±3.45)% and(30.11±5.93)%,respectively.The protein levels of CD97EGF and CD97stalk in MGC803 cells decreased by(26.25±5.73)% and(16.22±3.23)%,respectively.No change of the cell viability after siRNA transfection was observed.The cell number of migration and invasion in AGS cells was decreased by(67.63±12.03)% and(68.02±15.63)%,respectively.The cell number of migration and invasion in MGC803 cells was decreased by(14.92±2.03)% and(22.09±5.43)%,respectively.CONCLUSION: The siRNA effectively inhibits CD97 expression and restrains the migration and invasion capacities of gastric carcinoma cell lines,suggesting that CD97 plays an important role in the metastasis of gastric cancer.
【Fund】: 浙江省科技计划资助项目(No.2004C34010)
【CateGory Index】: R735.2
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