c-jun基因沉默对放射抗拒鼻咽癌裸鼠移植瘤生长及血管生成的影响

Silencing of c-jun inhibits growth and angiogenesis of radioresistant human nasopharyngeal carcinoma tumors implanted in nude mice

目的探讨c-jun基因沉默对人鼻咽癌放射抗拒CNE-2R细胞裸鼠移植瘤生长及血管生成的影响。方法用携带shRAN慢病毒载体感染CNE-2R细胞以沉默c-jun基因,构建c-jun-shRNA-CNE-2R细胞组(c-jun-shRNA组)以及阴性对照NC-shRNA-CNE-2R细胞组(NC组),以CNE-2R细胞为空白对照组,采用Western blot检测3组细胞中血管内皮生长因子(vascular endothelial growth factor,VEGF)蛋白的表达水平。构建裸鼠模型,观察3组细胞裸鼠皮下种植瘤生长曲线及体积;采用免疫组化法检测移植瘤组织中VEGF、CD34的表达,并检测组织内微血管密度(microvessel density,MVD)。结果 Western blot结果显示,c-jun-shRNA组细胞VEGF蛋白表达较NC组及空白对照组明显下调(P<0.05)。裸鼠模型实验中,c-jun-shRNA组移植瘤体积为(720.85±72.10)mm^3,质量为(0.42±0.04)g,MVD为11.69±3.30;NC组移植瘤体积为(1 196.81±90.69)mm^3,质量为(0.80±0.08)g,MVD为32.56±7.33;空白对照组移植瘤体积为(1 203.76±100.42)mm^3,质量为(0.83±0.05)g,MVD为35.45±4.87;c-jun-shRNA组种植瘤体积、质量、MVD较NC组和空白对照组低(P<0.05)。免疫组化检测结果显示,移植瘤组织中VEGF蛋白表达c-jun-shRNA组呈弱阳性,NC组及空白对照组组均呈强阳性。结论 c-jun基因沉默可抑制放射抗拒性鼻咽癌移植瘤生长及血管生成。

Objective To investigate the effect of c-jun silencing on the growth and angiogenesis of radioresistant human nasopharyngeal carcinoma CNE-2R cells implanted in nude mice. Methods Cells were infected with lentivirus expressing c-jun-targeting shRNA in CNE-2R cells （c-jun-shRNA group） or with lentivirus expressing negative control shRNA （NC group）;control cells were left uninfected. Levels of vascular endothelial growth factor （VEGF） in the three types of cells were measured using Western blotting. Positive clones were selected and transplanted in nude mice. Levels of CD34 and VEGF were analyzed using immunochemistry. Results VEGF protein levels were significantly lower in the c-jun-shRNA group than in the NC and control groups in vitro and in vivo. In the c-jun-shRNA group, tumor volume was（720.85±72.10） mms, tumor weight was （0.42±0.04） g, and MVD was 11.69±3.30. The corresponding results in the NC group were （1,196.81±90.69）mms, （0.80±0.08） g,and 32.56±7.33 ;the corresponding results in the control group were （ 1,203.76 ± 100.42 ） mm3, （ 0.83 ±0.05 ） g, and 35.45 ±4.87. Tumor volume, tumor weight, MVD and grading differed significantly between the c-jun-shRNA group and the other groups （P〈0.05）. Conclusion Sileneing the e-jun gene can inhibit the growth and angiogenesis of radioresistant nasopharyngeal carcinoma.