摘要
目的:探讨HSV-TK+GFP/GCV自杀基因系统对小鼠胰腺癌细胞系MPC体外及体内杀伤作用及其产生的旁观者效应.方法:通过RT-PCR从基因组文库中扩增出HSV-TK基因全长CDS序列,并将其与GFP基因定向克隆到质粒表达载体pcDNA3.1(+),构建重组质粒pcDNA3.1+/HSV-TK+GFP.脂质体法将重组质粒转染小鼠胰腺癌细胞株MPC细胞,得到带有HSV-TK和GFP基因的MPC/HSV-TK+GFP细胞,并将其分别用于体外和体内实验.结果:重组质粒pcDNA3.1+/HSV-TK+GFP导入小鼠胰腺癌细胞株MPC细胞.体外实验结果显示,当MPC/HSV-TK+GFP细胞数占混合细胞10%时,低浓度(20mg/L)的GCV就可将50%左右的肿瘤细胞杀死.体内实验结果显示GCV可明显抑制MPC/HSV-TK+GFP细胞在昆明小鼠体内的肿瘤形成.结论:HSV-TK和GFP基因转入小鼠胰腺癌细胞株MPC细胞并获得稳定表达,HSV-TK+GFP/GCV自杀基因系统在体内外对小鼠胰腺癌均有杀伤作用,且存在明显的旁观者效应.
AIM： To study in vitro therapeutic effect on mouse pancreatic cancer, as well as the bystander effect with HSV-TK suicide gene in combination with prodrug GCV. METHODS： HSV-TK and GFP were inserted into pcDNA3.1 （＋） to construct pcDNA3.1＋/ HSV-TK＋GFP, and pcDNA3.1＋/HSV-TK＋GFP was transferred into mouse pancreatic cancer cell MPC by Lipofectin. We then added GCV to these gene-modified cells and studied the sensitivity of the cells to GCV as well as the bystander effect. RESULTS： The gene modified pancreatic cancer cells MPC/HSV-TK＋GFP weresuccessfully developed. In vitro experiments showed that when the MPC/HSV- TK＋GFP cells accounted for 10% of hybrid cells, the low concentration （20 mg/L） of GCV was about 50% of tumor cell killing. In vivo results showed that the low concentration of GCV killed the cells. And tumor growth of the mouse model was inhibited. CONCLUSION： Our data demonstrate MPC/ HSV-TK＋GFP cells are sensitive to the treatment of GCV compared with unmodified tumor cells, and remarkable bystander effect is seen.
出处
《世界华人消化杂志》
CAS
北大核心
2009年第15期1498-1503,共6页
World Chinese Journal of Digestology
关键词
自杀基因治疗系统
旁观者效应
胰腺癌
小鼠
Suicide gene therapy system
Bystander effect
Pancreatic cancer
Mouse
