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作 者:童强[1] 郜元军[1] 李胜保[1] 张卫国[1] 王强[1]
机构地区:[1]湖北医药学院附属太和医院消化内科,湖北十堰442000
出 处:《临床内科杂志》2014年第1期57-59,共3页Journal of Clinical Internal Medicine
摘 要:目的 构建人CXCR4 mRNA的shRNA真核表达载体质粒,特异性抑制人食管癌细胞CXCR4的表达,并筛选抑制效果理想的质粒,研究对食管癌细胞迁移的影响.方法 以人CXCR4 mRNA编码区中3条不同序列作为RNA干扰靶点,分别构建3个shRNA真核表达载体质粒GYJ-1、GYJ-2、GYJ-3,并进行测序.用脂质体转染人食管癌细胞株Eca109,实时定量逆转录-聚合酶链反应(RT-PCR)检测mRNA水平.应用细胞划痕试验研究干扰质粒对食管癌细胞迁移的影响.结果 GYJ-1呈高效特异地抑制人CXCR4的表达;而GYJ-2及GYJ-3抑制效果较GYJ-1差.GYJ-1显著抑制食管癌细胞的迁移.结论 通过构建CXCR4的shRNA真核表达载体导入食管癌细胞,可有效抑制人食管癌细胞中CXCR4的表达,并影响食管癌细胞的迁移.Objective To specific suppress the expression of human CXCR4 gene in esophageal carcinoma by constructing the eukaryotic expression plasmid of shRNA, and to study the effct on cell mi- gration of esophageal carcinoma. Methods Plasmids containing three different sequences of human CXCR4 mRNA coding region were constructed. We gained three plasmids GYJ-1, GYJ-2 and GYJ-3. Plas- mids were identified by sequencing. Detect the effect of inhibition by real-time RT-PCR after transfecting human esophageal carcinoma cell line Ecal09 with liposomes. Detect cell migration by scratch test. Re- suits The expression of human CXCR4 gene was suppressed specific and effectively by GYJ-1. Yet GYJ- 2 and GYJ-3 were not the same. Cell migration of Eca-109 cell was also inhibit by GYJ-1. Conclusion CXCR4 expression in human esophageal carcinoma cells can be inhibited significantly by construction of eukaryotic expression vector expressing the shRNA, and cell migration of Eca-109 cell also can be inhibi- ted.
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