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Improvement of Human AFP Promoter Efficiency by CMV Early Enhancer
Ye Jingjia1, Chen Ping2, Jia Zhenyu3*, Li Chunchun1, Chen Lihong1, Cao Jiang1*
1Clinical Research Center, The 2nd Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310009, China;
2Institute of Clinical Research, Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, Hangzhou 3
2Institute of Clinical Research, Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, Hangzhou 3
Abstract: CMV early enhancer can be used to improve the efficiency of promoters of other genes. The impact of CMV early enhancer on the efficiency and specificity of AFP promoter determines whether it is suitable for AFP promoter in targeted gene therapy of hepatocyte carcinoma. In this study， human AFP enhancer/promoter and CMV early enhancer were cloned by PCR， and luciferase reporter vectors were constructed and transfected into hepatocyte carcinoma Hep3B， HepG2 and SMMC7721 cells， cervical carcinoma HeLa cells and breast cancer Bcap37 cells， with pGL4.74 vector as transfection control. Dual luciferase reporter assay was performed to analyze the efficiency of AFP promoter in these cells. The results showed that CMV early enhancer can markedly improve the efficiency of AFP enhancer/promoter in hepatocyte carcinoma cells (33.07-， 134.22- and 465.18-fold in Hep3B，HepG2 and SMMC7721 cells respectively)， and can also improve the efficiency of AFP enhancer/promoter in nonhepatocyte cells significantly (335.73- and 1096.81-fold in HeLa and Bcap37 cells respectively). Therefore， the CMV early enhancer improves the efficiency of AFP enhancer/promoter without cell-specificity. Side-effects may arise when applying CMV enhancer directly in AFP-targeted gene therapy of hepatocyte carcinoma due to nonspecific expression of therapeutic genes in normal tissues.
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