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

WU Xu-dong1,CHEN Wei-chang1,ZHU Shu-yang2,YE Jian-xin1 1.Department of Gastroenterology,First affiliated Hospital of Soochow University,Suzhou 215006,P.R.China2.Department of Pediatrics,North Courtyard of First People's Hospital of Chenzhou City,Chenzhou 423000,P.R.China  
OBJECTIVE:To investigate the effect of short hairpin RNA(shRNA)targeting at peroxisome proliferator-activated receptor gamma(PPARγ)gene on angiogenesis of hepatocellular carcinoma and its mechanicm in nude mice.METHODS:shRNA targeting at PPARγ gene was constructed and transfected into HCCLM3 cells(pshPPARγ group),meanwhile,the cells transfected 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)cm3,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)cm3,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,P0.01.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.
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