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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 peroxisome proliferator - activated receptor γ (PPARγ) gene silencing on intratumor angiogenesis in nude mice with hepatocellular carcinoma. Methods: The short hairpin RNA (PPARγ) expression plasmid was constructed and transfected into HCCLM3 cells (pshPPARγ group). The empty plasmid was transfected into the control group. The growth curve and volume of lung cancer and the number and grade of lung metastasis were observed. The expression of anti-CD34, vascular endothelial growth factor (VEGF) and thrombospondin (TSP1) were detected by immunohistochemistry. The expressions of MMP-2 and TIMP2 were detected by RT-PCR. Results: The volume of implanted tumor in pshPPARγ group was (1.86 ± 0.65) cm 3, the microvessel density (MVD) was 20.84 ± 6.38, and the number of lung metastases was (37.2 ± 0.7) / lung. The volume of control group was (4.86 ± 1.15) cm3, MVD was 39.48 ± 9.01, the number of lung metastases was (107.8 ± 6.1) / lungs, and the grading was mostly grade 3 to 4. The differences of tumor volume, MVD, Statistically significant, t values were 5.082,8.441 and 21.83, P <0.05. The expression of TSP1 protein in pshPPARγgroup was weakly positive or negative while the expression of MMP-2mRNA was down-regulated while the expression of TIMP2mRNA was down-regulated in the pshPPARγgroup (t = 11.34,8.44, P < 0.01. Conclusion: PPARγ gene silencing can reduce angiogenesis in implanted hepatocellular carcinoma, which may be related to the regulation of MMP-2 / TIMP2 expression and VEGF / TSP1 balance.