融合基因pCEAcd-tk前体药物体系的体外旁观者效应的差异性.docVIP

???? 融合基因pCEAcd-tk/前体药物体系的体外旁观者效应的差异性 ???? ???? 摘　要　目的： 探讨融合自杀基因pCEAcd-tk在导入到肺癌细胞后，其体外旁观者效应的细胞差异性。方法： 融合基因pCEAcd-tk提取后，脂质体介导的方法转染到6种肺癌细胞株(SPCA-1，GLC-82，A549，SC，HLAMP，L-78)，用MTT法测定在细胞转染前后对前体药物5-氟胞嘧啶（5-FC）和丙氧鸟苷（GCV）的敏感性和在体外的旁观者效应。结果： ① 脂质体介导的基因转染效率很低。② 在6种已转染的肺癌细胞株中，有2种肺癌细胞株（SPCA-1，A549）对前体药物5-FC和GCV有较高的敏感性；③ 旁观者效应显示，2种细胞株（SPCA-1，SC）有强的体外旁观者效应，1种细胞株(A549)有较强的旁观者效应，而另外3种细胞株（GLC-82，HLAMP和L-78）的旁观者效应较差。结论： 不同的细胞在转染了融合自杀基因以后，其体外旁观者效应存在较大的差异。关键词　自杀基因； 旁观者效应； 基因治疗； 5-氟胞嘧啶； 丙氧鸟苷《中国书资料分类法》分类号　Q344.13 Variation of Bystander Effect of E coli. CD/HSV1-tk and Prodrugs System in vitro Guo Yubin, Wu Xinyao, Luo Chaoquan， Guo Junming, Xu Daosong, Liu Min (Department of Biochemistry, Sun Yat-sen University of Medical Sciences, Guangzhou 510080) Abstract　Objective: To investigate the variation in vitro bystander effect among human lung cancer cell lines after transfection of pCEAcd-tk. Methods: pCEAcd-tk was extracted and transfected to six human lung cancer cell lines. Their sensitivity to 5-fluorocytosine (5-FC) and ganciclovir (GCV) was measured by MTT assay. Their bystander effect was also determined. Results: 1. The efficiency of lipid mediated gene transfer was very low; 2. Two out of six human lung cancer cell lines(SPCA-1，A549) have a high sensitivity to 5-FC and GCV; 3.Bystander effect: Two out of six human cancer cell lines（SPCA-1，SC） have a good bystander effect, and one out of them（A549）has a intermediate bystander effect, but three out of them （GLC-82，HLAMP and L-78）have a poor bystander effect. Conclusion： There are a variation in vitro bystander effect in six human lung cancer cell lines after transfected with pCEAcd-tk.Key words　suicide gene; bystander effect; gene therapy; 5-fluorocytosine; ganciclovir 　 自杀基因/前体药物体系治疗恶性肿瘤有一个很大的优势就是存在旁观者效应，不仅转染了自杀基因的肿瘤细胞在运用前体药物后被杀死，且没有被转染的肿瘤细胞也被杀死。这种旁观者效应显著地扩大了自杀基因杀伤肿瘤细胞的能力，在一定程度上弥补了当前基因转染效率低的问题。Sturtz［1］报道转染了胸苷激酶（tk）基因的7种神经胶质瘤细胞对前体药物GCV的敏感性不一样，那么旁观者效应是否存在细胞差异性？本研究选用我们前期构建的融合自杀基因pCEAcd-tk的表达体系转染到6种肺癌细胞株，对此进行了研究，现报道如下。1　材料与方法1．1