摘要
目的研究蜂毒素基因转染对肝癌细胞生物学性状的影响。方法将蜂毒素基因置于甲胎蛋白(AFP)转录调控序列驱动下,用细菌内高效同源重组法将目的基因重组于腺病毒质粒中;将重组腺病毒质粒用PacⅠ酶切线性化后,脂质体介导转染293细胞进行腺病毒的包装。携有蜂毒素基因的腺病毒感染肝癌细胞后,RT-PCR实验观察蜂毒素基因是否可以被转录,流式细胞术检测CD54表达情况。结果携蜂毒素基因的重组腺病毒载体构建成功;RT-PCR实验表明蜂毒素基因可以得到转录;流式细胞术结果表明,蜂毒素基因转染可以抑制CD54分子的表达。结论蜂毒素基因转染肝癌细胞后,可改变肿瘤细胞的生物学行为,使其恶性度降低。
Objective To observe the effects of melittin gene transfection on the biological behavior of hepatocarcinoma cells. Methods Melittin gene was recombined into a shuttle plasmid under the control by α-fetoprotein (AFP) transcription regulatory element (rAFP), and the gene of interest was then recombined into an adenoviral backbone plasmid through a simplified efficient bacterial homologous recombination system. The recombinant adenoviral plasmids were subsequently linearized with PacⅠand transfected into 293 cells mediated by lipofectin to generate the desired recombinant adenoviruses. After infections of the hepatocarcinoma cells with the resultant viruses carrying melittin gene was achieved, melittin gene transcription was verified by way of RT-PCR, and CD54 expression was measured by flow cytometry. Results Recombinant adenoviral vectors con- taining melittin gene were successfully constructed, and melittin gene transcription was approved by RT-PCR. Flow cytometry results showed that melittin gene transfection inhibited CD54 expression in the hepatocarcinoma cells. Conclusion Melittin gene transfection may result in changes of the biological behavior of hepatocarcinoma cells, thus decreasing the malignancy of the tumor cells.
出处
《第一军医大学学报》
CSCD
北大核心
2003年第4期300-305,共6页
Journal of First Military Medical University
基金
军队"九五"重点基金(98ZD3269)
上海市科技发展基金(98XD140200)
上海市百人计划资助项目(97BR044)~~
关键词
蜂毒素
基因转染
腺病毒感染
肝癌
甲胎蛋白
转录调控序列
melittin gene
alpha-fetoprotein
transcription regulatory elements
adenovirus vector
CD54
