Caspase-3 gene transfected with LIGHT gene:: can it be used for therapy of human hepatocellular carcinoma?
Caspase-3 gene transfected with LIGHT gene:: can it be used for therapy of human hepatocellular carcinoma?
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DOI：
10.1515/cclm.2008.094
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发表时间：
2008-01-01
影响因子：
6.8
通讯作者：
Han, Bing
中科院分区：
文献类型：
--
作者：
Lu, Yun;Wu, Li-Qun;Han, Bing
背景：本研究旨在检测转染HepG(2)细胞中凋亡因子caspase-3的表达情况，为基因方法治疗原发性肝癌提供可行的评价。方法：从大肠杆菌JM-109中提取pcDNA4C-LIGHT cDNA；然后，通过阳离子脂质体介导的方法将pcDNA4C-LIGHT cDNA转入HepG2细胞中。同时设立空白组作为对照组，并于转染后12 h、24 h、48 h、3 d、5 d收集HepG(2)细胞。通过ELISA鉴定上清液中caspase-3的表达。为所测定的一组样品生成标准曲线。 SPSS分析差异有统计学意义。结果：Caspase-3蛋白量在48h时最多，第5天最少。对照组中caspase-3的分泌量没有增加。相关系数为0.9986，具有显着性意义。结论：pcDNA4C-LIGHT通过脂质体介导有效转染人HepG(2)细胞。转染组中caspase-3的表达增加。该研究为基因方法治疗肝癌提供了必要的理论支持。
Background: The aim of this study was to detect the expression of apoptosis factor caspase-3 in transferred HepG(2) cells and provide feasible evaluation of the treatment for primary liver cancer with gene methods.Methods: The pcDNA4C-LIGHT cDNA was extracted from Escherichia coli JM-109; then, the pcDNA4C-LIGHT cDNA was transferred into the HepG2 cells by a cationic liposome mediated method. Meanwhile, the blank group was established as the control group and the HepG(2) cells were collected after transfection at 12 h, 24 h, 48 h, 3 days and 5 days. The expression of caspase-3 was identified in the supernatants by ELISA. A standard curve was generated for the set of samples assayed. Statistical significance was analyzed by SPSS.Results: The quantity of caspase-3 protein was the greatest at 48 h and the least on day 5. The secretion of caspase-3 did not increase in the control group. The coefficient of correlation was equal to 0.9986 and had evident significance.Conclusions: The pcDNA4C-LIGHT was effectively transfected in human HepG(2) cells mediated by liposome. The expression of caspase-3 increased in the transfected group. This study provides necessary theoretic support for the treatment of liver cancer with gene methods.