Baculovirus vector-mediated transfer of sodium iodide symporter and plasminogen kringle 5 genes for tumor radioiodide therapy.
Baculovirus vector-mediated transfer of sodium iodide symporter and plasminogen kringle 5 genes for tumor radioiodide therapy.
复制标题
DOI：
10.1371/journal.pone.0092326
复制
发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Li B
中科院分区：
文献类型：
--
作者：
Zhang M;Guo R;Shi S;Miao Y;Zhang Y;Li B
在设计癌症治疗方案时，肿瘤细胞及其支持的内皮细胞都应考虑靶向细胞杀伤。本文研究了杆状病毒介导的碘化钠同转运体（NIS）和纤溶酶原kringle 5 （K5）编码基因转移后，放射性碘和抗血管生成治疗联合应用的可行性。构建了一种由人类端粒酶逆转录酶（hTERT）启动子和早期生长反应1 （Egr1）启动子驱动的K5基因控制的含有NIS基因的重组杆状病毒。双荧光素酶报告基因试验证实了hTERT转录的激活。Western blot和Real-Time PCR检测NIS和K5基因的表达。在杆状病毒感染的Hela细胞中，通过摄取125I和131I的细胞毒性证实了NIS的功能性活性。采用流式细胞术分析131i诱导的K5对杆状病毒感染的人脐静脉内皮细胞（HUVECs）的凋亡作用。在体内进行NIS报告基因成像和131I的治疗实验。最后用CD31免疫染色法测定治疗后肿瘤微血管密度（MVD）。hTERT转录的激活在肿瘤细胞中特异性上调。在杆状病毒感染的HeLa细胞中，NIS基因表达明显增加，而在MRC5细胞中则没有。在NaClO4的抑制下，Hela细胞对125I的摄取明显增加，131I处理显著降低了细胞存活率。131I诱导的K5基因表达呈剂量和时间依赖性升高，导致huvec细胞凋亡。此外，131I SPECT成像清楚地显示重组杆状病毒感染的宫颈肿瘤异种移植物。治疗后，肿瘤生长明显迟缓。CD31免疫染色证实MVD明显降低。重组杆状病毒通过靶向肿瘤及其支持血管，支持了基于nis的碘化碘治疗与基于k5的抗血管生成治疗相结合的有前景的策略。
Both tumor cells and their supporting endothelial cells should be considered for targeted cell killing when designing cancer treatments. Here we investigated the feasibility of combining radioiodide and antiangiogenic therapies after baculovirus-mediated transfer of genes encoding the sodium iodide symporter (NIS) and plasminogen kringle 5 (K5). A recombinant baculovirus containing the NIS gene under control of the human telomerase reverse transcriptase (hTERT) promoter and the K5 gene driven by the early growth response 1 (Egr1) promoter was developed. Dual-luciferase reporter assay was performed to confirm the activation of hTERT transcription. NIS and K5 gene expression were identified by Western blot and Real-Time PCR. Functional NIS activity in baculovirus-infected Hela cells was confirmed by the uptake of 125I and cytotoxicity of 131I. The apoptotic effect of 131I-induced K5 on baculovirus-infected human umbilical vein endothelial cells (HUVECs) was analyzed by a flow cytometry-based assay. In vivo, NIS reporter gene imaging and therapeutic experiments with 131I were performed. Finally, the microvessel density (MVD) in tumors after treatment was determined by CD31 immunostaining. The activation of hTERT transcription was specifically up-regulated in tumor cells. NIS gene expression markedly increased in baculovirus-infected HeLa cells, but not in MRC5 cells. The Hela cells showed a significant increase of 125I uptake, which was inhibited by NaClO4, and a notably decreased cell survival rate by 131I treatment. Expression of the K5 gene induced by 131I was elevated in a dose- and time-dependent manner and resulted in the apoptosis of HUVECs. Furthermore, 131I SPECT imaging clearly showed cervical tumor xenografts infected with recombinant baculovirus. Following therapy, tumor growth was significantly retarded. CD31 immunostaining confirmed a significant decrease of MVD. The recombinant baculovirus supports a promising strategy of NIS-based raidoiodide therapy combined with K5-based antiangiogenic therapy by targeting both the tumor and its supporting vessels.
登录
查看更多内容
DOI：
10.1016/j.mrfmmm.2011.11.002
发表时间：
2012-02-01
影响因子：
2.3
作者：
Podlevsky, Joshua D.;Chen, Julian J. -L.
通讯作者：
Chen, Julian J. -L.
影响因子：
5.1
作者：
Kawashita, Y;Ohtsuru, A;Yamashita, S
通讯作者：
Yamashita, S
丁酸钠增强A549肺腺癌细胞中杆状病毒介导的钠/碘同向转运蛋白基因的表达
影响因子：
1.5
作者：
Guo, Rui;Zhang, Yifan;Li, Biao
通讯作者：
Li, Biao
影响因子：
5.8
作者：
Cengic, N;Baker, CH;Spitzweg, C
通讯作者：
Spitzweg, C
影响因子：
4.8
作者：
Cao, YH;Chen, A;Llinas, M
通讯作者：
Llinas, M