目的:构建兔VEGF反义cDNA真核表达载体(pcDNA3.1-ASVEGF);利用反义核酸技术调控脑脊液源性转移的形成,MRI活体动态监测,探讨抗血管生成治疗在脑脊液源性转移形成和预后中的价值。方法:构建兔VEGF反义核酸真核表达质粒;VX2细胞接种后不同时间行MRI检查。使用GE公司的SIGNA1.5T磁共振成像系统,膝关节正交线圈。行常规及DCE-MRI检查,分析时间-信号曲线参数SLE值。在每次MR检查前抽取实验兔血液和脑脊液标本做VEGF的ELISA检测。所有实验兔在最后一次检查完毕后取材行病理检查和VEGF免疫组化染色分析。结果:经酶切及测序鉴定,成功获得兔子反义VEGF核酸真核表达载体(pcDNA3.1-ASVEGF);通过对DCE-MRI参数SLE、肿瘤生长速度、动物存活时间的分析,各组之间有统计学意义;DCE-MRI参数SLE与VEGF的IHS评分以及与血液及脑脊液VEGF表达均呈正相关(P〈0.05)。结论:VEGF反义核酸对脑脊液源性转移瘤具有调控作用。MRI是一种可靠、无创的脑脊液源性转移病变反义核酸治疗监测工具。
Objective: Using molecular biologlcal technique to construct cukaryotic expression vector of rabbit VEGF antisense cDNA;Using MRI to monitor the regulation for leptomeningeal metastasis. Methods: Eukaryotic expression vector of rabbit VEGF antisense eDNA was constructed; MRI examinations were performed at different times after inoculation on a SIGNA 1.5 tesla scanner. The knee coil was used for imaging. Routine and DCE-MRI were acquired. SLE of time-signal curve, the VEGF level and pathology were analysed. Resuits: We successfully constructed eukaryotic expression vector of rabbit VEGF antisense eDNA (pcDNA3.1-ASVEGF). SLE was differnet in different groups after different days inoculating the VX2 tumor cells.The numbers and size of foci showed difference between group A, B,C and group D,E ( P 〈 0.01). VEGF immunohistochemical score and the VEGF level in the plasma and CSF bore a positive correla- tion with SLE ( P 〈 0.01 ) by rank correlation statistical method ( P 〈 0.05). Conclusion: Anti,use VEGF eDNA can inhibit the VEGF expression of leptoraeningeal loci on molecule level. MRI can accurately show the leptomeningeal metastases, which can be used in early diagnosis and monitoring therapeutic effectiveness on leptomeningeal metastasis.