肺癌基因治疗新思路:肺癌细胞血管内皮生长因子表达的抑制

New way to treat lung cancer gene:inhibition of vascular endothelial growth factor expression in lung cancer cell

目的:探讨硫修饰和脂质体包裹的反义寡核苷酸(ASODN)抑制Lewis肺癌细胞血管内皮生长因子(vascularendothelialgrowthfactor,VEGF)基因表达的效应,减少肺切除导致的呼吸功能衰竭发生的可能性。方法:将经脂质体包裹并部分硫代修饰后的VEGFASODN,正义寡核苷酸(SODN),错义寡核苷酸(MODN)加入培养的Lewis肺癌细胞中,采用免疫组织化学SP法及RT-PCR技术检测肺癌细胞中VEGF蛋白及VEGFmRNA表达,同时测定各上清液刺激血管内皮细胞生长的受抑情况。结果:VEGFASODN能明显抑制Lewis肺癌细胞VEGF蛋白及VEGFm-RNA的表达,而且可以通过抑制VEGF的表达抑制内皮细胞的生长,并对内皮细胞的抑制作用存在浓度依赖性,5,10,20μmol/L浓度组的内皮细胞生长抑制率分别约为13.77%,48.85%,76.29%。VEGFSODN和VEGFMODN却无此作用。结论:VEGFASODN抑制肺癌细胞表达VEGF,进而抑制内皮细胞生长,可以成为肺癌基因治疗的一种新的策略。

AIM:To investigate inhibition of the vascular endothelial growth factor(VEGF) expression of Lewis' lung cancer cell by the phosphorothioate modify and the liposome encapsulation antisense oligodeoxynucleotides(ASODN),and reduce the possibility of weakening respiratory function induced by cutting the lung. METHODS:The phosphorothioate modify and the liposome encapsulation VEGF ASODN,VEGF SODN and VEGF missense oligodeoxynucleotides(MODN) were added into the medium of Lewis' lung cancer cell respectively.Expression of VEGF protein and VEGFmRNA were detected by streptavidin perosidase(SP) immunohistochemical method and RT PCR.And the inhibitory effects of vascular endothelial cell proliferation stimulated by supernatants were determined. RESULTS:VEGF ASODN can significantly inhibit the expression of VEGF protein and VEGFmRNA of Lewis' lung cancer cell and the proliferation of endothelial cells in concentration dependent fashion. The inhibitory rate of proliferation of endothelial cells in 5,10,20 μmol/L group were about 13.77%, 48.85%, 76.29%respectively,however,there was no such effect between VEGF SODN and VEGF MODN. CONCLUSION:VEGF ASODN may inhibit the expression of VEGF,even the proliferation of endothelial cell,it is hopeful that VEGF ASODN become a new antigiogenesis therapy strategy in lung carcinoma.