摘要
背景:肝细胞癌(HCC)是全球最常见的恶性肿瘤之一,目前手术、介入、放化疗等常规治疗效果均不理想,开发有效、毒副作用相对较小的新药成为研究者关注的重点。目的:观察东亚钳蝎镇痛抗肿瘤肽(AGAP)基因真核表达载体对HCC细胞的靶向表达和毒性作用。方法:真核表达质粒pAFP-AGAP经酶切和测序鉴定后转染甲胎蛋白(AFP)阳性人HCC细胞株HepG2以及AFP阴性人宫颈癌细胞株HeLa和正常人肝细胞株L02。逆转录聚合酶链反应(RTPCR)检测AGAP mRNA表达,CCK-8方法检测细胞毒性作用。结果:各组细胞转染质粒pAFP-AGAP后,AGAPmRNA表达仅见于AFP阳性HepG2细胞,而AFP阴性HeLa细胞和L02细胞均无AGAP mRNA表达。HepG2细胞转染质粒pAFP-AGAP 48 h后形态发生明显改变,细胞生长显著受抑(P<0.01),48 h和72 h时抑制率分别为44.4%和74.6%,而HeLa细胞和L02细胞的形态和生长均未受明显影响。结论:AGAP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HCC靶向基因治疗的有力工具。
Background： Hepatocellular carcinoma （HCC） is one of the most common malignancies worldwide; however, the efficacy of conventional therapies such as surgical operation, interventional therapy and radiochemotherapy is not so satisfactory. New treatment modalities with higher efficacy and lower toxic reaction should be pursued. Aims： To assess the targeting and cytotoxie effects of eukaryotic expression vector carrying analgesic-antitumor peptide （AGAP） gene derived from Buthus martensii Karseh on HCC cells. Methods： Eukal-yotic expression plasmid pAFP-AGAP was identified by enzyme digestion and sequencing, and then transfeeted into α-fetoprotein （AFP）-positive human HCC cell line HepG2 and AFP-negative human cervical carcinoma cell line HeLa and normal human hepatic cell line LO2. The expression level of AGAP mRNA was determined by reverse transcriptase polymerase chain reaction （RT-PCR）, and Cell Counting Kit-8 was used to analyze the eytotoxicity. Results： AGAP mRNA was detected in AFP-producing HepG2 cells but not in HeLa and LO2 cells after plasmid pAFP-AGAP transfection. Forty-eight hours after transfection with plasmid pAFP-AGAP, significant morphological changes and growth inhibition （P〈0.01） were observed in HepG2 cells, the inhibition rate at 48- hour and 72-hour was 44.4% and 74.6%, respectively; whereas no such changes were found in HeLa and LO2 cells. Conclusions： AGAP gene eukaryotic expression vector can selectively target the HCC cells with high cytotoxic efficacy, which might be a promising effective tool for HCC gene therapy.
出处
《胃肠病学》
2009年第12期742-745,共4页
Chinese Journal of Gastroenterology
关键词
镇痛抗肿瘤肽
蝎毒素
癌
肝细胞
基因疗法
Analgesic-Antitumor Peptide
Charybdotoxin
Carcinoma, Hepatocellular
Gene Therapy
