电转染血管能抑素基因对C_(57)小鼠Lewis肺癌生长和转移的影响

Effects of Canstatin gene Transfected by Electroporation on Growth and Metastasis of Lewis Lung Carcinoma in C57 Mice

目的建立Lewis肺癌移植瘤模型,人血管能抑素(canstatin)基因重组表达载体电穿孔法转染至肿瘤局部,观察对肿瘤生长和转移的抑制作用。方法将Lewis肺癌细胞接种于C57BL小鼠皮下,成瘤后将30只小鼠随机分成3组,分别瘤体内注射血管能抑素重组载体、空载体和生理盐水,基因枪于注射部位电击,2周后PT-PCR法检测肿瘤组织中血管能抑素基因的表达并观察小鼠体重、肿瘤体积变化及肺转移结节数目。结果成功建立Lewis肺癌皮下移植瘤模型;电击2周后重组载体组仍存在血管能抑素基因的表达,重组载体组小鼠肿瘤体积由(1.03±0.11)cm3增加至(1.51±0.19)cm3,而空载体组由(1.01±0.09)cm3增至(2.45±0.18)cm3、NS组由(1.02±0.10)cm3增至和(2.54±0.21)cm3;重组载体组体积与后两组比较(P<0.01),即生长速度受到显著的抑制。重组载体组、空载体组和NS组的肺转移结节数分别为(3.50±1.24),(7.60±2.51)和(7.70±2.45);重组载体组与后两组比较(P<0.01),即肿瘤转移亦受到显著的抑制。结论血管能抑素可明显抑制Lewis肺癌移植瘤的生长与转移。

Objectives To establish animal model of Lewis lung carcinoma in C57 mice and study the inhibitory effect of canstatin gene transfected by electroporation on the growth and metastasis of Lewis lung carcinoma. Methods Lewis lung carcinoma cells were subcutaneously inoculated in CATBL mice to make animal model of tumor. Thirty mice in whom the transplanted tumor in the mice up to 1cm^3grew were randomly divided into 3 groups,where the eukaryotic expression vector of pCMV-Script, the recombinant pCMV-Script/Canstatin vector and normal saline were injected into tumor tissue respectively. The injective areas were electroporated by particle gun in the mice. The expression of canstatin mRNA in the tumor tissues was detected by RT-PCR, the size and weight of the subcutaneous tumors were observed and the number of metastasis node in the lungs was calculated 14 days after the electroporation treatment. Results The animal model of tumor was successfully established. Two weeks later, the expression of canstatin mRNA in the tumor tissues of the recombinant vector group could detected by RT-PCR, the tumor sizes in the recombinant vector group slowly grew from（ 1.03 ± 0. 11 ） cm^3 to （ 1.51 ± 0. 19 ） cm^3 , naked vector group from （ 1.01 ±0.09 ） cm^3 to（2. 45±0. 18）cm^3 and normal saline group from（ 1.02±0.10）cm^3 to（2.54±0.21 ）cm^3. The number of metastasis node was 3.50±1. 24, 7.60± 2.51 and 7.70 ±2.45 respectively in the recombinant vector group, naked vector group and normal saline group. There was significant difference in the volume of the tumor and the number of metastasis node in the lungs between the recombinant vector group and the naked vector group or normal saline group（ P 〈0.01 ）. Conclusion Canstatin has strong inhibitory effect on the growth and metastasis of Lewis lung carcinoma in mice.