E-cadherin启动子调控的慢病毒荧光报告载体的构建

Construction of Lentiviral-based System with Fluorescent Reporter Gene Regulated by E-cadherin Promoter

E钙黏素(E-cadherin)的缺失是上皮-间质转化(EMT)的重要标志,而EMT可促进肿瘤细胞的侵润及转移。为进一步研究E-cadherin与EMT的关系,研究构建了E-cadherin启动子驱动的黄色荧光蛋白慢病毒表达载体,构建的载体经酶切鉴定正确;采用磷酸钙法包装出慢病毒,取病毒上清感染人肝癌PLC/PRF/5细胞株,通过荧光观察和qRT-PCR实验结果显示E-cadherin在靶细胞中稳定高表达。该载体的构建为深入研究E-cadherin相关的EMT现象提供了良好的工具。

The loss of E-cadherin is a significant marker of epithelial-mesenchymal transition （EMT）, which promotes the progression of tumor invasion and metastasis. To further examine the relationship be- tween E-eadherin and EMT, the lentiviral vector with yellow fluorescent protein reporter gene driven by E- cadherin promoter is successfully constructed. The supernatant of virus-producing cells transfected by cal- cium phosphate is then used to transfect PLC/PRF/5 cells. The fluorescent microscopy and qRT-PCR re- sults show that the E-cadherin gene is expressed highly and stably in the target cells. This work is expec- ted to provide a high-quality transfection vector/or further research on the relevant functions of the E-cad- herin gene and EMT.