高强度聚焦超声治疗对肝癌细胞裸鼠移植瘤低氧诱导因子表达和细胞凋亡的影响  

Effect of HIFU on hypoxia-inducible factor( HIF-1α) expression and cell apoptosis in liver cancer cell xenografts of nude mice

作　　者：付志豪[1] 武伦 乔正荣[2] 周世骥[3] 李生伟[1] 

机构地区：[1]重庆医科大学附属第二医院肝胆外科,重庆400010 [2]重庆市第五人民医院普外科,重庆400062 [3]重庆医科大学附属第二医院胃肠外科,重庆400010

出　　处：《基础医学与临床》2014年第5期648-653,共6页Basic and Clinical Medicine

基　　金：国家自然科学基金(81301975;81272570);重庆市卫生局重点资助项目(渝卫2012-1-040;渝卫2010-1-68)

摘　　要：目的 探讨高强度聚焦超声（HIFU）治疗后的残余肝癌组织中低氧诱导因子（HIF-1α）、P53和凋亡的变化及关系.方法 建立肝癌HpG2细胞系裸鼠移植瘤模型30只,采用CZF-Ⅱ型HIFU治疗仪治疗,随机分为对照组、治疗后1、3及5d和1及2周组.HE染色观察标本病理变化;TUNEL法检测残余癌组织凋亡及计算凋亡指数;免疫组化、Western blot和实时荧光定量PCR技术检测HIF-1α、P53蛋白和mRNA水平.结果 治疗后有残余肿瘤细胞和大片坏死区域.与其他组相比,凋亡指数在3d组显著升高（P＜0.05）,在5d、1和2周组逐渐下降.HIF-1α和P53蛋白在各组中出现强弱不同的表达.HIF-1α和P53蛋白和mRNA在治疗后1～3d表达逐渐升高,在3d组显著高于其他组（P＜0.05）,在5d、1和2周组逐渐下降.结论 HIFU治疗肝癌细胞裸鼠移植瘤后可以促进残余癌细胞凋亡,此可能与HIF-1α、P53基因表达异常关系密切.Objective To study the effects of hypoxia-inducible factor （HIF-1α） on apoptosis of the residual tumor cells in nude mice after treatment by high-intensity focused ultrasound （HIFU）.Methods Human HCC implantation was established in 30 nude mice.CZF-Ⅱ HIFU therapeutic apparatus was used for treatment.Nude mice were randomly divided into control group and treatment group （included 1 d group,3 d group,5 d group,1 week group,and 2 weeks group）.Pathological changes were observed with HE staining;Cell apoptosis was detected by DNA agarose gel electrophoresis and terminal deoxynucleotidyltransferase-mediated Dutp nick end-labeling （TUNEL） assay; SP immunohistochemistry was used to detect HIF-1α protein;Western blot and real-time quantitative PCR were used to detect HIF-1α,P53 protein and mRNA expression levels.Results HE showed that there were residual tumor cells and large necrotic areas after treatment.Compared with other groups,the apoptosis index in 3 d group was significantly higher （P ＜ 0.05）,while in the 5 d group,1 week group,2 weeks group gradually decreased.Immunohistochemistry showed that the expression of HIF-1α,P53 protein was different in each group.Compared with other groups,Western blot and RT-PCR showed an increase of HIF-1 α,P53 protein and mRNA levels in groups after treatment of 1 to 3 days,and peaked in 3 d group（P ＜ 0.05）.Conclusions HIFU treatment can induce consistent apoptosis in residual tumor,which may be related with disordered expression of HIF-1 α and P53.

关 键 词：肝癌 低氧诱导因子 凋亡 高强度聚焦超声 

分 类 号：R735.7[医药卫生—肿瘤]