Zhu Mice, Yang Jian-ru, Jiang Yue-quan, Chen Shi-feng, Fu Xin-ping, Tian Ze-dan
Department of Cardiothoracic Surgery, the Second Affiliated Hospital of Chongqing Medical University, Chongqing 400010, China.
Sichuan Da Xue Xue Bao Yi Xue Ban. 2011 Mar;42(2):152-6.
To observe the killing effect of recombinant type I herpes simplex virus (HSV-I) with Gibbon ape leukemia virus membrane fusion glycoprotein (GALV.fus) gene on lung adenocarcinoma in vitro and in vivo.
Recombinant HSV-I plasmids (HSV-UL38P-GALV.fus, HSV-CMVP-GALV.fus, HSV-CMVP-EGFP) was introduced into green monkey kidney cells (Vero) by liposome to amplify the virus, which were propagated in Vero cells and purified by cesium chloride density purification, titrated by TCID50 method. The three recombinant viruses were named as Synco-2, Synco-1 and Baco-1 respectively, and were transfected into lung adenocarcinoma cell line A549 cell and human lung adenocarcinoma xenografts which were established in nude mice subcutaneously to observe the expression and transfection of recombinant plasmids; mice model was divided to A (Control) group, B (Baco-1) group, C (Synco-1) group, D (Synco-2) group and E (Synco-2) group. The antitumor and cytotoxic effects of the virus in vitro or in vivo were investigated simultaneously.
Recombinated HSV-I virus were packed successfully, the titre of Baco-1, Synco-1 and Synco-2 were 3 x 10(10) pfu/mL, 1X 10(11) pfu/mL and 4 x 10(10) pfu/mL respectively. The virus produced clear antitumor effects in vitro, the oncolytic activity of Synco-2 and Synco-1 was superior to that of Baco-1 (P < 0.01). The striking antitumor effect was seen when the virus was given subcutaneously in established xenografts in the animals. Tumor volume in Group C and D decreased significantly compared those in Group A and B (P < 0.01). The same result was observed in tumour weight (P < 0.01), and we also find that there was statistical significance between Group C and D in tumour quality at last two weeks (P < 0.01).
The three recombinant HSV-I were packaged, amplificated and purified successfully. Recombinant GALV. fus gene system controlled by special promoter and mediated by available carrier has potent activity against lung cancer both in vitro or in vivo, and maybe a new promising candidate for investigative gene therapy of this malignancy.
观察携带长臂猿白血病病毒膜融合糖蛋白（GALV.fus）基因的重组I型单纯疱疹病毒（HSV-I）对肺腺癌的体内外杀伤作用。
采用脂质体法将重组HSV-I质粒（HSV-UL38P-GALV.fus、HSV-CMVP-GALV.fus、HSV-CMVP-EGFP）导入绿猴肾细胞（Vero）中扩增病毒，在Vero细胞中传代培养后经氯化铯密度梯度离心法纯化，采用TCID50法进行病毒滴度测定。将三种重组病毒分别命名为Synco-2、Synco-1和Baco-1，转染肺腺癌细胞系A549细胞及皮下接种于裸鼠建立的人肺腺癌移植瘤，观察重组质粒的表达及转染情况；将小鼠模型分为A（对照）组、B（Baco-1）组、C（Synco-1）组、D（Synco-2）组和E（Synco-2）组，同时观察病毒在体内外的抗肿瘤及细胞毒性作用。
重组HSV-I病毒包装成功，Baco-1、Synco-1和Synco-2的病毒滴度分别为3×10(10)pfu/mL、1×10(11)pfu/mL和4×10(10)pfu/mL。病毒在体外产生明显的抗肿瘤作用，Synco-2和Synco-1的溶瘤活性优于Baco-1（P<0.01）。在动物已建立的移植瘤中皮下注射病毒时观察到显著的抗肿瘤作用。C组和D组的肿瘤体积与A组和B组相比显著减小（P<0.01）。肿瘤重量也观察到相同结果（P<0.01），并且我们还发现最后两周C组和D组在肿瘤质量方面存在统计学差异（P<0.01）。
三种重组HSV-I成功包装、扩增及纯化。由特定启动子控制并由有效载体介导的重组GALV.fus基因系统在体内外对肺癌均具有强大的活性，可能是这种恶性肿瘤研究性基因治疗的一种新的有前景的候选方案。
