Combinatorial anti-angiogenic gene therapy in a human malignant mesothelioma model
Combinatorial anti-angiogenic gene therapy in a human malignant mesothelioma model
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DOI：
10.3892/or.2015.4058
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发表时间：
2015-08-01
期刊：
影响因子：
4.2
通讯作者：
Kasahara, Noriyuki
中科院分区：
文献类型：
--
作者：
Kubo, Shuji;Takagi-Kimura, Misato;Kasahara, Noriyuki
抗血管生成基因治疗是一种很有前途的癌症治疗策略，然而，它很少在恶性间皮瘤中进行测试，恶性间皮瘤是一种与石棉相关的高度侵袭性肿瘤，预后不良。在本研究中，我们调查是否抗血管生成因子，如血管抑素，内皮抑素和可溶性形式的血管内皮生长因子受体2（sFlk 1）能够抑制内皮细胞增殖，通过慢病毒介导的基因转移到恶性间皮瘤细胞培养。我们还评估了双药策略是否具有更大的治疗益处。使用慢病毒载体转导人恶性胸膜间皮瘤MSTO-211 H细胞，所述慢病毒载体单独表达血管抑制素、内皮抑制素和sFlk 1，并通过内部核糖体进入位点连接到增强的绿色荧光蛋白（EGFP）标记基因表达。单独表达EGFP的慢病毒用作对照。通过蛋白质印迹分析和荧光显微镜证实所得到的命名为MST 0-A、MST 0-E、MST 0-F和MST 0-C的细胞稳定表达相应的蛋白质。在这些细胞中的任何细胞之间的体外生长速率中未观察到差异。然而，与MSTO-C相比，MSTO-A、MSTO-E和MSTO-F的共培养在体外显示出对人脐静脉内皮细胞生长的显著抑制。此外，MST 0-A、MST 0-E和MST 0-F中的任何两种的组合显著增强功效。这些结果表明，针对内皮生长因子信号传导不同途径的组合抗血管生成基因疗法有可能比单药方案获得更大的治疗效果。
Anti-angiogenic gene therapy represents a promising strategy for cancer; however, it has rarely been tested in malignant mesothelioma, a highly aggressive tumor associated with asbestos with poor prognosis. In the present study, we investigated whether anti-angiogenic factors such as angiostatin, endostatin and the soluble form of vascular endothelial growth factor receptor 2 (sFlk1) were able to inhibit endothelial cell proliferation via lentivirus-mediated gene transfer into malignant mesothelioma cells in culture. We also assessed whether a dual-agent strategy had greater therapeutic benefit. Human malignant pleural mesothelioma MSTO-211H cells were transduced using lentiviral vectors that individually expressed angiostatin, endostatin and sFlk1 and linked to enhanced green fluorescent protein (EGFP) marker gene expression via an internal ribosome entry site. The lentivirus expressing EGFP alone was used as a control. The resultant cells designated as MSTO-A, MSTO-E, MSTO-F and MSTO-C were confirmed by western blot analysis and fluorescence microscopy to stably express the corresponding proteins. No differences were observed in the in vitro growth rates between any of these cells. However, co-culture of MSTO-A, MSTO-E and MSTO-F showed significant suppression of human umbilical endothelial cell growth in vitro compared with that of MSTO-C. Furthermore, a combination of any two among MSTO-A, MSTO-E and MSTO-F significantly enhanced efficacy. These results suggest that combinatorial anti-angiogenic gene therapy targeting different pathways of endothelial growth factor signaling has the potential for greater therapeutic efficacy than that of a single-agent regimen.