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

发布时间：2018-05-11 03:38

本文选题：血管内皮生长因子A + 高强度聚焦超声　；参考：《重庆医学》2017年06期

【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(P0.05)。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。
[Abstract]:Objective to observe the effect of high intensity focused ultrasound (HIFU) on the treatment of nude mice liver transplantation tumor, and to investigate the therapeutic mechanism of HIFU after injection of HSV-TKcarrying herpes simplex virus-thymidine kinase gene (HSV-TK). Methods 60 nude mice models of subcutaneous transplanted tumor were established by using human hepatoma HepG2 cells. After treatment with CZF type HIFU machine, 60 nude mice were treated. Two hundred 渭 L microbubble HSV-TK phosphatidylinositol proteoglycan (PIP-3) was injected into the tail vein of rats. The microbubble HSV-TK(B group was injected with HSV-TK GPC3(C group, and the microbubble GPC3(E group was treated with HSV-TKN D group. The microbubble GPC3(E group was injected 3 times every 3 days. After each injection, according to the ultrasonic monitoring of the arrival of microbubbles to target tissues, the DZC type ultrasound transfection apparatus was used to irradiate the Agna group with 2W / cm ~ 2 ~ (-1) MHz ~ (2 +) for 5 min. 48 hours after the first injection, each group was injected intraperitoneally with 0.2mL ganciclovir (100mg / kg 路kg ~ (-1) 路d ~ (-1) and continuously injected for 14 days. After treatment, the levels of microvessel density (MVD) were measured by immunohistochemical method, and the changes of hypoxia inducible factor-1 伪 (HIF-1 伪) and vascular endothelial growth factor (VEGF) were measured by Western blot and immunohistochemistry. Q-PCR was used to detect the transcription level of HIF-1 伪 -VEGF gene in tumor. Results after 14 days of treatment, the expression of HIF-1 伪 -VEGF protein and the level of HIF-1 伪 mRNA in tumor tissue of group A were significantly higher than that of group A (P 0.05). Conclusion the treatment of HSV-TK with nano-ultrasound molecular probe can reduce the level of VEGF HIF-1 伪 and MVD in residual tumor tissue after HIFU treatment, thus inhibit the growth of tumor and improve the therapeutic effect of HIFU.
【作者单位】：四川省德阳市人民医院肝胆外科;重庆医科大学附属第二医院肝胆外科;
【基金】：国家自然科学基金资助项目(81272570)
【分类号】：R735.7

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