The targeted gene (KDRP-CD/TK) therapy of breast cancer mediated by SonoVue and ultrasound irradiation in vitro
The targeted gene (KDRP-CD/TK) therapy of breast cancer mediated by SonoVue and ultrasound irradiation in vitro
复制标题
SonoVue联合超声照射介导的乳腺癌靶向基因（KDRP-CD/TK）治疗
DOI：
10.1016/j.ultras.2011.08.002
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发表时间：
2012-01-01
期刊：
影响因子：
4.2
通讯作者：
Zhang Ping
中科院分区：
文献类型：
--
作者：
Li Xing-hua;Zhou Ping;Zhang Ping
自杀基因治疗已成为乳腺癌的有效治疗方法，而超声靶向微泡破坏（UTMD）已成为基因治疗领域的热门话题。在本研究中，使用UTMD用重组质粒pEGFP-KDRP-CD/TK转染具有KDR启动子的MCF-7细胞和没有KDR启动子的LS174T细胞。重组质粒pEGFP-KDRP-CD/TK成功转染MCF-7和LS174T细胞，转染效率无显着差异(p > 0.05)。 RT-PCR显示CD/TK融合基因在MCF-7细胞中表达，但在LS174T细胞中不表达。在细胞毒性实验中，转基因MCF-7细胞对前药5-FC和GCV敏感。当同时施用 5-FC 和 GCV 时，细胞抑制率显着高于仅施用其中一种前药时所达到的细胞抑制率 (p < 0.001)。此外，施用5-FC、GCV以及5-FC和GCV两者所达到的抑制率均显着高于基因转染率21.92+/-3.64%（p<0.001）。然而，转基因LS174T细胞对任何前药都不敏感。这些结果表明UTMD是一种安全、有效、有针对性的基因传递系统。并且KDR启动子能够驱动MCF-7细胞中CD/TK双自杀基因靶点的表达，并且KDRP-CD/TK基因对MCF-7细胞的体外靶向杀伤作用与CD/TK融合基因的表达具有良好的协同作用。 (C) 2011 Elsevier B.V. 保留所有权利。
Suicide gene therapy has become an effective therapy for breast cancer, and ultrasound targeted microbubble destruction (UTMD) has become a popular topic in the gene therapy field. In this study, MCF-7 cells with the KDR promoter and LSl74T cells without the KDR promoter were transfected with the recombinant plasmid pEGFP-KDRP-CD/TK using UTMD. The recombinant plasmid pEGFP-KDRP-CD/TK was transfected into MCF-7 and LS174T cells successfully with no significant difference in transfection efficiency (p > 0.05). By RT-PCR, the CD/TK fusion gene was shown to be expressed in MCF-7 cells but not expressed in LS174T cells. In a cytotoxicity experiment, transgenic MCF-7 cells were sensitive to the prodrugs 5-FC and GCV. When both 5-FC and GCV were administered, the rate of cellular inhibition was significantly greater than that achieved when only one of the prodrugs was administered (p < 0.001). Moreover, the inhibition rates achieved administering 5-FC, GCV and both 5-FC and GCV were all significantly greater than the gene transfection rate of 21.92 +/- 3.64% (p < 0.001). However, transgenic LS174T cells were not sensitive to any prodrug. These results demonstrated that UTMD is a safe, effective and targeted gene delivery system. Also, the KDR promoter can drive expression of the CD/TK double suicide gene target in MCF-7 cells, and the targeted killing effect of the KDRP-CD/TK gene on MCF-7 cells in vitro has good synergy with expression of the CD/TK fusion gene. (C) 2011 Elsevier B.V. All rights reserved.