假单胞菌外毒素体外靶向抗肝细胞癌的实验研究

中国临床药理学与治疗学 ›› 2010, Vol. 15 ›› Issue (4): 372-375.

假单胞菌外毒素体外靶向抗肝细胞癌的实验研究

刘扬, 吴金明, 金思思, 申苏建, 吴利敏, 江宏峰, 金颖

温州医学院附属第一医院消化内科,温州 325000,浙江

收稿日期:2010-02-01 修回日期:2010-03-01 发布日期:2020-09-17
通讯作者: 吴金明,男,博士,教授,研究方向:肝脏病学。 E-mail:phdwu0003@yahoo.com.cn
作者简介:刘扬,男,硕士研究生,研究方向:肝脏病学。Tel: 13695895734 E-mail: liuyangmd@163.com

Empirical study of Pseudomonas aeruginosa exotoxin A targeting on hepatocellular carcinoma in vitro

LIU Yang, WU Jin-ming, JIN Si-si, SHEN Su-jian, WU Li-min, JIANG Hong-feng, JIN Yin

Department of Gastroenterology, the First Affiliated Hospital of Wenzhou Medical College, Wenzhou 325000, Zhejiang,China

Received:2010-02-01 Revised:2010-03-01 Published:2020-09-17

摘要/Abstract

摘要： 目的: 构建携铜绿假单胞菌外毒素PE38基因及甲胎蛋白(AFP)启动子的重组载体,并研究其在体外对AFP阳性肝细胞癌(hepatocellular carcinoma, HCC)的靶向杀伤作用。方法: 构建重组免疫毒素表达质粒pAFP-PE38,通过转染细胞观察其作用,RT-PCR法测定PE38 mRNA表达,CCK-8法检测细胞毒性。结果: 各组细胞转染质粒pAFP-PE38后仅AFP阳性的HepG2细胞表达PE38 mRNA,且形态发生改变,生长显著受抑(P＜0.01),48 h 和 72 h 抑制率分别为 27.2%、58.3%,而AFP阴性的PC-3和HeLa细胞未受明显影响。结论: PE38基因真核表达载体可实现HCC基因治疗的靶向性和高效性,有望成为肝细胞癌靶向基因治疗的有力工具。

关键词: PE38, 甲胎蛋白, 基因治疗

Abstract: AIM: To construct a gene-modified hepatocellular carcinoma (HCC) specific PE38 expression vector regulated by cis-acting element of AFP, and explore its anti-HCC effect in vitro. METHODS: Eukaryotic expression plasmid pAFP-PE38 was constructed and then transfected into different cell lines.The expression level of PE38 mRNA was detected by RT-PCR. Cytotoxicity was detected by cell counting kit-8 (CCK-8). RESULTS: PE38 mRNA was detected only in AFP-positive HepG2 cells after plasmid transfection. Significant morphological changes and growth inhibition (P＜0.01) were observed in HepG2 cells, the inhibition rate at 48 h and 72 h were 27.2% and 58.3%,respectively; whereas no such changes were found in AFP-negative PC-3 cells and HeLa cells. CONCLUSION: PE38 gene eukaryotic expression vector can selectively targets on HCC cells with high efficacy, which might be used as an effective tool for HCC gene therapy.

Key words: PE38, α-fetoprotein, Gene therapy

中图分类号:

R965.2

刘扬, 吴金明, 金思思, 申苏建, 吴利敏, 江宏峰, 金颖. 假单胞菌外毒素体外靶向抗肝细胞癌的实验研究[J]. 中国临床药理学与治疗学, 2010, 15(4): 372-375.

LIU Yang, WU Jin-ming, JIN Si-si, SHEN Su-jian, WU Li-min, JIANG Hong-feng, JIN Yin. Empirical study of Pseudomonas aeruginosa exotoxin A targeting on hepatocellular carcinoma in vitro[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2010, 15(4): 372-375.

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