摘要
目的 探讨经导管动脉化疗栓塞术(TACE)和微波消融(MWA)治疗VX2肝癌诱导的低氧诱导因子(HIF)-1α糖酵解与微血管密度相关性.方法 选取19只新西兰兔,建立18只VX2肝癌模型.随机分为对照组(建模不做处理,n=6)、TACE组(经肝动脉注射碘化油和吡柔比星乳剂,n=6)、MWA组(经皮穿刺肿瘤后行MWA,n=6).肿瘤长至1.5～2 cm时行相应手术治疗.术后2周处死实验兔,取肿瘤及瘤周些许肝组织.苏木精-伊红(HE)染色观察肿瘤细胞形态.免疫组化检测肿瘤HIF-1α、微血管密度(CD34、CD105标记)、血管内皮细胞生长因子(VEGF)、Ki67表达.蛋白质印迹实验检测糖酵解关键酶己糖激酶(HK)2、丙酮酸激酶(PK)M2、乳酸脱氢酶(LDH)A、葡萄糖转运体(GLUT)1、GLUT4及丙酮酸脱氢酶激酶(PDK)1蛋白表达.结果 18只兔均建模成功,手术成功率为100％.HE染色显示肿瘤细胞排列紊乱.免疫组化检测显示,TACE组、MWA组与对照组相比,HIF-1α、CD34、CD105、VEGF及Ki67表达升高(均P＜0.05);蛋白印迹实验分析显示,与对照组相比,TACE组、MWA组术后GLUT1、GLUT4表达升高(均P＜0.05);PKM2、HK2表达量MWA组升高(均P＜0.05),但TACE组的差异无统计学意义(均P＞0.05);LDHA、PDK1表达量TACE组升高(均P＜0.05),但MWA组的差异无统计学意义(均P＞0.05).Pearson相关性分析结果表明,肿瘤内微血管密度与糖酵解代谢呈正相关.结论 TACE和MWA治疗后VX2肝癌模型糖酵解代谢上调,且与肿瘤血管新生正相关.
Abstract
Objective To explore the correlation between HIF-1α regulation of glycolysis and microvessel density induced by transcatheter arterial chemoembolization(TACE)and microwave ablation(MWA)in experimental rabbits with VX2 hepatocellular carcinoma(HCC).Methods Nineteen New Zealand rabbits were selected,and VX2 HCC model was successfully established in 18 rabbits.The 18 rabbits with VX2 HCC were randomly divided into control group(no any treatment was given after modeling,n=6)and TACE group(receiving TACE with iodized oil and pirobicin emulsion,n=6),and MWA group(receiving MWA,n=6).When the tumor of experimental rabbits grew to 1.5-2 cm size,the corresponding treatment started.The experimental rabbits were sacrificed two weeks after the treatment,the tumor and a few peritumoral liver tissues were removed and sent for laboratory testing.Tumor cell morphology was examined by hematoxylin-eosin(HE)staining.Immunohistochemical assay was used to detect the hypoxia-inducing factor 1α(HIF-1α),microvascular density(CD34,CD105 markers),vascular endothelial growth factor(VEGF),and Ki67 expression.Western blot was used to detect the expressions of key glycolytic enzymes hexokinase 2(HK2),pyruvate dehydrogenase 2(PKM2),lactate dehydrogenase(LDHA),glucose transporter(GLUT 1 and GLUT4),and pyruvate dehydrogenase kinase 1(PDK1).The correlation between glycolysis and tumor microvascular density was evaluated by Pearson correlation analysis.Results A total of 18 rabbits were successfully modeled,the surgical success rate was 100％.HE staining examination demonstrated that the arrangement of the tumor cells was disorganized.Immunohistochemical assay indicated that in TACE group and MWA group the expressions of HIF-1α,CD34,CD105,VEGF,and Ki67 were remarkably elevated when compared with the control group(all P＜0.05).Western blot analysis revealed that when compared with the control group in TACE group and MWA group the expressions of GLUT1 and GLUT4 were obviously elevated(both P＜0.05);the expressions of PKM2 and HK2 were higher in MWA group(both P＜0.05),but no significant difference in TACE group(both P＞0.05);and the expressions of LDHA and PDK1 were higher in TACE group(both P＜0.05),but no significant difference in MWA group(both P＞0.05).Pearson correlation analysis showed that a positive correlation existed between the microvessel density and glycolytic metabolism in the tumor.Conclusion After receiving TACE and MWA treatment,in rabbit VX2 HCC models the glycolytic metabolism becomes up-regulated,which bears a positive relationship to tumor angiogenesis.(J Intervent Radiol,2024,33:390-396)
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