摘要
目的:探讨血管内皮生长因子C(VEGF-C)基因沉默对裸鼠移植乳腺癌淋巴管和血管生成的影响。方法:构建VEGF-C小分子干扰RNA(VEGF-C/SiRNA)慢病毒载体,转染靶细胞MCF-7,Real-time PCR检测VEGF-C敲减效率,将其接种于裸小鼠背部皮下(KD组),并设置MCF-7细胞对照组(CON组)及空载体对照组(NC组),观察裸小鼠的成瘤情况,通过免疫组化检测VEGF-C表达、微血管密度(MVD)及微淋巴管密度(LVD)。结果:VEGF-C/SiRNA转染MCF-7细胞后,与对照组相比,VEGF-C表达显著降低,敲减效率为49.2%,P=0.017。接种后各组细胞均能成瘤,KD组6只,NC组7只,CON组8只,成瘤情况差异无统计学意义,P=0.631。HE染色结果显示,KD组可见肿瘤细胞明显减少,出现较多凋亡和坏死细胞。免疫组化检测显示,CON组VEGF-C蛋白表达(+)1只,(++)1只,(+++)4只;NC组表达(++)1只,(+++)5只;KD组表达(+)5只,(++)1只,KD组移植瘤VEGF-C蛋白表达显著低于对照组,P=0.02。肿瘤组织内LVD计数,KD组5.33±1.63,CON组8.50±1.05,NC组8.17±1.83,KD组显著低于NC组和CON组,差异有统计学意义,P=0.005;MVD计数,KD组8.83±1.72,CON组11.02±1.41,NC组11.17±2.48,KD组MVD略低于CON组及NC组,但差异无统计学意义,P=0.097。结论:慢病毒介导的VEGF-C/siRNA可有效敲减VEGF-C表达,显著减少乳腺癌组织的淋巴管生成,但对血管生成无显著影响。
OBJECTIVE：To establish the nude mouse model with breast cancer MCF-7 cells in which VEGF-C was interfered with Lentivirus-Mediated Small Interfering RNA（VEGF-C/siRNA）, and investigate the relationship between VEGF-C expression and microvessel density （MVD） and lymphatic microvessel density （LVD）. METHODS： VEGF-C/ siRNA vectors were constructed and transducted into MCF-7 breast cancer cells,and real-time PCR was performed to detect the mRNA expression of VEGF-C. The MCF-7 cells which transfected with siRNA Vectors were injected into the nude mouse,and the volume of tumor was measured. The VEGF-C expression, MVD and LVD in transplant tumors were detected by immunohistochemistry. RESULTS： VEGF-C gene expression was down-regulated significantly in VEGF-C/ siRNA interfered MCF-7 ceils by 49.2% （P=0. 017）. The transplant tumor could grow in 3 groups of nude mice,6 in KD group,7 in NC group,8 in CON group. Cell number of transplant tumor in KD group decreased,and apoptosis and necrosis were observed in many cancer cells with HE staining. The immunohistochemical results showed that VEGF-C protein expression in KD group was decreased significantly（CON group： 1＋, 1＋＋, 4＋＋＋;NC group. 1 ＋＋, 5 ＋＋＋ ;KD group： 5＋ ,1＋＋ ,P=0.02）. LVD were 5.33土1.63,8.17土1.83,and 8.50土1.05 in KD group,NC group and CON group,re- spectively. LVD in KD group was lower significantly（P=0. 005）. There was not difference of MVD in KD group（8.83土1.72） ,NC group （11.17土2.48） and CON group （11; 02土1.41,P=0. 097）. CONCLUSION： The VEGF-C expression can be successfully knocked down with VEGF-C/siRNA Lentivirus, VEGF-C can stimulate tumor lymphangiogenesis and have no effect on tumor angiogenesis.
出处
《中华肿瘤防治杂志》
CAS
北大核心
2013年第8期583-587,共5页
Chinese Journal of Cancer Prevention and Treatment
关键词
乳腺肿瘤
血管内皮生长因子C
微血管密度
微淋巴管密度
裸
小鼠
breast cancer
vascular endothelial growth factor-C
microvessel density
lymphatic vessel density
nude mice
