HIFU联合载HSV-TK基因的纳米级超声分子探针治疗对裸鼠肝移植瘤血管生成的影响  

The effect of HIFU combined with nanoscale ultrasound molecular probes with simple virus thymidine kinase gene on angiogenesis in the nude mouse tumor

作　　者：谢辉[1] 刘山[1] 曾辉[1] 李友伟[1] 许进[1] 丁雄[2] 刘长安[2] 李峰[1] 

机构地区：[1]四川省德阳市人民医院肝胆外科,618000 [2]重庆医科大学附属第二医院肝胆外科,400010

出　　处：《重庆医学》2017年第6期725-728,共4页Chongqing medicine

基　　金：国家自然科学基金资助项目(81272570)

摘　　要：目的观察高强度聚焦超声(HIFU)治疗裸鼠肝移植瘤后,再注入载单纯疱疹病毒-胸苷激酶基因(HSV-TK)的纳米级超声分子探针治疗效果,并探讨其治疗机制。方法用人肝癌HepG2细胞构建裸鼠皮下移植瘤模型60只,采用CZF型HIFU机治疗后,分别经鼠尾静脉注射200μL的微泡+HSV-TK+磷脂酰肌醇蛋白多糖-3(GPC3)(A组)、微泡+HSV-TK(B组)、HSV-TK+GPC3(C组)、HSV-TK(D组)、微泡+GPC3(E组)、PBS液(F组),每3天1次,共注射3次。每次注射后,根据超声监测微泡到达靶组织的情况,对A、B、D、E组用DZC型超声转染仪给予2W/cm^2、1MHz、5min超声辐照。首次注射48h后,各组经腹腔注射0.2mL更昔洛韦(GCV,100mg·kg^(-1)·d^(-1)),连续注射14d。治疗结束后,用免疫组织化学法测残瘤中微血管密度(MVD)水平,用Western blot和免疫组织化学法测量残癌中缺氧诱导因子-1α(HIF-1α)和血管内皮生长因子(VEGF)的变化情况,用Q-PCR检测肿瘤中HIF-1α、VEGF基因的转录水平。结果治疗14d后,B、C、D、E、F组肿瘤组织中HIF-1α、VEGF蛋白表达量,VEGF及HIF-1αmRNA水平,MVD均明显高于A组(P<0.05)。结论载HSV-TK的纳米级超声分子探针治疗可降低HIFU治疗后残瘤组织中的VEGF、HIF-1α及MVD水平,从而抑制肿瘤的生长,提高HIFU的治疗效果。Objective To observe the change of the protein and gene expression of hypoxia inducing factor-1α（HIF-1α）and vascular endothelial growth factor（VEGF）in the nude mouse tumor,which has been treated by HIFU combined with nanoscale ultrasound molecular probes with HSV1-TK gene microvascular density.Methods Sixty nude mice were implanted with HepG2 Cells to establish subcutaneous transplanted tumor.Divided this mice into six groups at random after treated by HIFU：MB＋HSV-TK＋GPC3（group A）,MB＋HSV-TK（group B）,HSV-TK＋GPC3（group C）,HSV-TK（group D）,MB＋GPC3（group E）,PBS（group F）.They were injected into the tail vein every after 3days.Mice in group A,B,D and E were exposed to ultrasound by 2W/cm2,1 MHz,5mintues and 0.2mL ganciclovi（GCV）was intraperitoneally injected at the first 48 hours after injection.After the treatment,immunohistoche were used to detect the microvascular density（MVD）,Western blot and immunohistoche was employed to test the protein change of the VEGF and HIF-1α,Q-PCR was used to test the mRNA gene transcription of VEGF and HIF-1αin the tumor tissues.Results After 14 days,the protein expression of HIF-1αand VEGF in group A was significantly lower than that in group B,C,D,E and F（P〈0.05）,the MVD level in the tumor is also like this,and the difference is statistically significant.Conclusion Anoscale ultrasound molecular probes with HSV1-TK can reduce the the level of VEGF,MVD and HIF-1αin the tumor which has been treated by HIFU,so it can inhibit tumor growth and improve the therapeutic efficacy after HIFU treatment.

关 键 词：血管内皮生长因子A 高强度聚焦超声 缺氧诱导因子1 分子探针 微血管密度 

分 类 号：R735.7[医药卫生—肿瘤]