| The study of expression and correlation between HMF-1α and VEGF in hepatocellular carcinoma pre-and post-transcatheter arterial embolizationPurpose:To investigate the expression levels of serum hypoxia inducible factor-1 alpha(HIF-1α)and vascular endothelial growth factor(VEGF)pre-and post-transcatheter arterial embolization(TAE)in patients with hepatocellular carcinoma(HCC),and the correlations between prognosis factors and serum HIF-1α as well as VEGF levels.Materials and methods:Forty-two consecutive patients fulfilling criteria for HCC(experimental group)and six diagnosing Hepatic hemangioma patients(control group)undergoing TAE were from June 2014 to December 2015 enrolled into the study.According to the tumor location and size,the different doses of iodized oil and embolic agents were used in embolization The serum HIF-la and VEGF levels of HCC patient pre-TAE and 3 days,7 days,1 month post-TAE were analyzed using enzyme-linked immunosorbent assay(ELISA),and compared with that of 6 Hepatic hemangioma patients.Patients were divided into complete response(CR),partial response(PR),stable disease(SD)and progressive disease(PD)groups according to the therapeutic efficacy:Pearson correlation was used to analyze the correlation between different clinical factors and serum HIF-la and VEGF,and correlation between different HIF-1α and VEGF levels also evaluated.Results:The expression levels of serum HIF-1α and VEGF in HCC patients were 162.19±43.03 pg/mL and 278.70±55.55 pg/mL pre-TAE,and both of them were significantly higher than those in control group(27.14±5.94pg/mL and 54.51±11.38pg/mL,all P<0.05).Three days post-TAE,both serum HIF-1α(332.17±7.37pg/mL)and VEGF(398.37±69.38 pg/mL)reached the peak values,significantly higher than those pre-TAE(P<0.05).Seven days post-TAE,expression levels of them were decreased(245.20 ±84.52 pg/mL and 334.25±83.88pg/mL respectively),but still significantly higher than those before TAE(P<0.05).One month post-TAE,the levels of both HIF-1α(118.67±43.70pg/mL vs 252.07±67.17pg/mL)and VEGF(199.33±31.68pg/mL vs 354.75±92.82pg/mL)in CR group were significantly lower than those in PR+ SD+PD group(P<0.05).The level of serum HIF-1α was positively correlated with serum VEGF level(r=0.67,P<0.05).Both serum HIF-1α and VEGF levels were observed to be correlated with portal vein thrombi and:metastasis(All P<0.05).Conclusion:The serum HIF-la and VEGF might play an important role in the process and relapse of HCC.Meanwhile,hypoxia-induced angiogenesis plays a key role in the process of invasion and metastasis of HCC.They might be considered as predictors of the efficacy of TAE and patient survival condition.Liposomal curcumin inhibits hypoxia-induced angiogenesis after transcatheter arterial embolization in VX2 rabbit liver tumorsPurpose:the purpose of the study is to investigate the correlations between HIF-la as well as VEGF levels in VX2 rabbit liver tumors,and the inhibition of hypoxia-imduced angiogenesis after transcatheter arterial embolization by liposomal curcumin.Materials and methods:(1)Liposomal curcumin:Lipids and curcumin(SPC:CHOL:curcumin,20:1:2)were dissolved and mixed in dichloromethane,and then the solvent was removed to yield a film.The solution was transferred to containers and the frozen,and then the frozen lipid cake was placed on a vacuum pump and lyophilized until dried.Hydration of the dry film/cake was accomplished by adding phosphate-buffered saline solution to the container of dry film and agitating at a tenperature of 45℃.Lastly the particles were downsized by extrusion and stored.(2)VX2 rabbit tumor model:The tumor was harvested from the carrier rabbits and minced in 0.9%normal saline solution.The recipient rabbit were anesthetized and exposed through abdominal incision.The fresh tumor tissues measuring 1mm ×1mm × 1mm were embedded into the left liver lobe.All VX2 rabbits were given normal feeding for 18-20 days.(3)Experimental procedures:The VX2 rabbits were divided into three groups.Group 1(control group n=18)animals underwent sham embolization with 0.9%normal saline solution.Group 2(TAE-treated group n=18)animals underwent embolization with lipiodol and 90-180 um polyvinyl alcohol particles.Group 3(liposomal curcumin TAE-treated group n=18)animals underwent embolization with liposomal curcumin(20mg/kg body weight)mixed with lipiodol and 90-180 um PVA particles.Based on the time of sacrifice,the animals in each group were then divided into three subgroups:subgroup 1(n=6),6 hours after experimental intervention;subgroup 2(n=6),24 hours after experimental intervention;and subgroup 3(n=6),3 days after experimental intervention.(4)Interventional Procedures:All VX2 models were performed under digital subtraction angiography.The micro-catheter was advanced to super-select the common hepatic,proper hepatic,and tumor’s blood supply artery in sequence.For the control group,2 mL of 0.9%normal saline solution was injected.For the TAE-treated group,lipiodol(0.1 mL/kg body weight)was slowly administered and 0.1 mL of PVA particles was carefully injected.For the liposomal curcumin TAE-treated group,liposomal curcumin(20mg/kg body weight)mixed with lipiodol(0.1 mL/kg body weight)was slowly administered,and then,embolization was done with PVA particles.(5)After embolization,animals in each subgroup were sacrificial at 6 hours,24 hours,and 3 days,and the tumor samples were collected.Immunohistochemical staining was performed to evaluate expression of hypoxia-inducible factor-la(HIF-1α)and vascular endothelial growth factor(VEGF)proteins,and microvessel density(MVD).Real-time polymerase chain reaction was performed to examine VEGF mRNA levels.Results:(1)The average particle size of the liposomal curcumin was 118.2±0.91 nm,the zeta potential or total charge on the surface of the liposomes was-1.66±0.14 mV.Surface morphology of the liposomes presented a spheriacal shape in standard electronic modules.(2)In the control group,the size of the tumor revealed a trend of obvious growth after sham embolziaton.The tumor size slightly decreased in the liposomal curcumin TAE-treated group compared to the TAE-treated group,and the difference was not statistically significant on the third day after embolization(P>0.05).(3)In the TAE-treared group,the HIF-1α positive cells were highly presented at the residual tumors near necrosis of the tumor peripheral zone.In contrast,the levels of HIF-1α protein in the liposomal TAE-treated group significantly decreased compared to the TAE-treated group at three time points(P<0.05).(4)Repreentative VEGF-positive cells were highly presented in the residual tumors near necrosis zone in the TAE-treated group.The levels of VEGF mRNA and protein substantially decreased and had a statistical difference at three time points in the liposomal curcumin TAE-treated tumors compared with the TAE-treated tumors(P<0.05).(5)The mean MVD presented an obvious rising from 6 hours to 3 days in the TAE-treated group(P<0.05).The mean MVD had significantly decreased in the liposomal curcumin TAE-treated gourp compared with the TAE-treated group.(6)Spearman’s rank correlation analysis showed that HIF-1α protein significantly correlated with VEGF mRNA(r=0.705,P=0.001)and protein(r=0.655,P=0.003),and MVD(r=0.521,P=0.027).A significant correlation between VEGF protein and MVD was noted as well(r=0.519,P=0.027).Conclusion:Hypoxia-induced angiogenesis plays an important role in the process of invasion and relapse of liver tumor.Liposomal curcumin downregulates HIF-1α protein levels and inbibits hypoxia-induced angiogenesis after embolization in VX2 rabbit liver tumors. |
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