PPARγ基因沉默抑制裸鼠肝癌血管生成的初步研究  

Effect of RNA interference targeting at peroxisome proliferator-activated receptor gamma gene on angiogenesis of hepatocellular carcinoma in nude mice

作　　者：吴旭东[1] 陈卫昌[1] 朱树养[2] 叶建新[1] 

机构地区：[1]苏州大学附属第一医院消化内科,江苏苏州215006 [2]郴州市第一人民医院北院儿二区,湖南郴州423000

出　　处：《中华肿瘤防治杂志》2009年第7期497-501,共5页Chinese Journal of Cancer Prevention and Treatment

摘　　要：目的:探讨过氧化物酶体增殖物活化受体γ(PPARγ)基因沉默对肝癌裸鼠模型瘤内血管生成的影响。方法:构建PPARγ短发夹状RNA表达质粒并转染HCCLM3细胞(pshPPARγ组),以空质粒转染为对照组。观察两组皮下种植瘤生长曲线及体积,肺转移灶数量及分级。免疫组织化学法检测抗-CD34、血管内皮生长因子(VEGF)、血小板反应蛋白(TSP1)表达;RT-PCR检测MMP-2和TIMP2表达。结果:pshPPARγ组种植瘤体积为(1.86±0.65)cm3,微血管密度(MVD)为20.84±6.38,肺转移灶数量为(37.2±0.7)个/肺,分级为1～2级,对照组体积为(4.86±1.15)cm3,MVD为39.48±9.01,肺转移灶数量为(107.8±6.1)个/肺,分级多为3～4级,两组种植瘤体积、MVD和转移灶数量、分级的差异有统计学意义,t值分别为5.082、8.441和21.83,P值均<0.05。pshPPARγ组TSP1蛋白阳性表达,而VEGF蛋白为弱阳性或阴性表达,MMP-2mRNA表达下调,而TIMP2mRNA表达下调,与对照组比较差异有统计学意义,t值分别为11.34、8.44,P值均<0.01。结论:PPARγ基因沉默可降低肝癌种植瘤血管生成,与调节MMP-2/TIMP2表达,影响VEGF/TSP1平衡有关。OBJECTIVE： To investigate the effect of short hairpin RNA （shRNA） targeting at peroxisome proliferator-activated receptor gamma （PPART） gene on angiogenesis of hepatocellular carcinoma and its meehanicm in nude mice. METHODS： shRNA targeting at PPARy gene was constructed and transfected into HCCLM3 cells （pshPPAR7 group）, meanwhile, the cells transfeeted with vector plasmids were as a control group. Positive clones were selected and the transplanted tumor animal models constructed in nude mice, and the growth and volume of tumors were observed. After 8 weeks, PPARγ mRNA and protein in transplanted tumor tissues were detected, and pulmonary metastases were counted and graded. Moreover, Anti-CD34, VEGF and TSP1 were stained by the immuocytochemistry method, and MMP-2 and TIMP2 mRNA were detected by RT-PCR. RESULTS： PPARγ mRNA was down regulated significantly in the pshPPARγ group, and its inhibition rate was 82%. In the pshPPARγ group, the tumor volume was （1.86±0. 65） cma , MVD was 20. 84± 6.38, and pulmonary metastases were at grade 1--2 and 37.2±0.7 per lung; while in the control group, tumor volume was （4.86 ± 1.15） cm^3 , MVD was 39.48±9.01, and pulmonary metastases were at grade 3- 4 and 107.8 ± 6.1 per lung, In the pshPPARγ group, TSP-1 protein presented positive, and VEGF protein presented weakly positive or negative. The expression of MMP-2 mRNA decreased, TIMP2 mRNA increased in the pshPPARγ group, and there were significant differences compared with the control group,t= 11.34 or 8. 44, P〈0. 01. CONCLUSION： RNA interference targeting at PPARγ gene can balance TSP1/VEGF, through regulating the expression of MMP- 2/TIMP2 to impair angiogenesis of HCC in nude mice.

关 键 词：短发夹状RNA 肝细胞癌 血管生成 过氧化物酶体增殖物活化受体Γ 

分 类 号：R735.7[医药卫生—肿瘤]