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1、抑制CD24表达对SW620细胞粘附能力影响的研究论文邓启亮王新颖王伟飞彭亮青海涛姜泊【摘要】目的:检测了DLD1、HT29、HepG2、STLYCD24mut细胞株表达CD24后,肿瘤细胞与纤维连接蛋白的结合能力显著增强。1材料与方法1.1材料1.1.1细胞株:HT29、HepG2、SI1640中,每周传代2次。siRNA干扰技术使用的是Invintrogen公司Lipofectamine2000tm试剂。具体步骤参照试剂盒。2.2细胞株的筛选2.2.1半定量RTPCR检测各株细胞CD24mRNA表达水平总RNA的提取按照英俊公司TRIZOLReagent步骤,各取5g总RNA
2、,按RevertAidTMHMinusFirstStrandcDNASynthesisKit操作说明合成cDNA扩增CD24和GAPDH采用相同的cDNA上样量(1L)和反应参数,反应参数设置:94℃预变性5min,然后进入以下36个循环:94℃变性45sec,55℃退火1min,72℃延伸1min。循环完毕,72℃延伸5min。2.2.2流式细胞检测SI1640中,调整细胞浓度为1×108/L;按200L/孔加入细胞悬液,37℃温育1h;取出96孔板,1×PBS冲洗3次,9每孔加入5mg/ml的MTT20μL和全培200μL;用酶联免疫检测仪在570nm处读取OD值。3结果3.1C
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