Intravesical administration of exogenous microRNA-145 as a therapy for mouse orthotopic human bladder cancer xenograft.
Intravesical administration of exogenous microRNA-145 as a therapy for mouse orthotopic human bladder cancer xenograft.
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DOI：
10.18632/oncotarget.4129
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发表时间：
2015-08-28
期刊：
影响因子：
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通讯作者：
Azuma H
中科院分区：
文献类型：
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作者：
Inamoto T;Taniguchi K;Takahara K;Iwatsuki A;Takai T;Komura K;Yoshikawa Y;Uchimoto T;Saito K;Tanda N;Kouno J;Minami K;Uehara H;Hirano H;Nomi H;Kiyama S;Akao Y;Azuma H
我们先前报道了microRNA（miR）-145在人膀胱癌细胞中的水平减弱。在本研究中，我们通过使用原位人膀胱癌异种移植模型研究了膀胱内施用miR-145是否可能是控制膀胱癌的潜在治疗策略。将miR-145转染253 J B-V细胞后，通过MTT、Western blotting、Hoechst 33342染色和体外伤口愈合试验检测miR-145异位表达的影响。此外，通过将B-V细胞植入小鼠膀胱壁中，建立了小鼠原位人膀胱癌模型。然后评估将miR-145膀胱内注射到这些小鼠中的抗癌作用。用miR-145转染253 J B-V细胞在体外诱导细胞凋亡和细胞迁移抑制。Western blotting结果显示，C-Myc、socs 7、FSCN 1、E-cadherin、β-catenin和catenin δ-1水平降低，PI 3 K/Akt和Erk 1/2信号通路代偿性增加。在体内，用miR-145处理的小鼠显示出76%的肿瘤生长抑制，动物存活显著延长（p = 0.0183 vs.对照）。Western blotting结果显示，细胞凋亡相关基因和细胞运动相关基因在体外均显著减少。此外，PI 3 k/Akt和Erk 1/2信号通路，这是在体外激活的补偿方式，在体内减少。因此得出结论，在该人膀胱癌异种移植模型中，膀胱内施用外源性miR-145是膀胱癌的有效疗法。
We previously reported that the level of microRNA (miR)-145 is attenuated in human bladder cancer cells. In this current study, we investigated whether intravesical administration of miR-145 could be a potential therapeutic strategy for controlling bladder cancer by using an orthotopic human bladder cancer xenograft model. Following transfection of 253J B-V cells with miR-145, the effects of the ectopic expression of miR-145 were examined by performing MTT, Western blotting analysis, Hoechst33342 staining, and wound healing assay in vitro. Also, a mouse orthotopic human bladder cancer model was established by inoculating 253J B-V cells into the bladder wall of mice. The anti-cancer effects of intravesical injections of miR-145 into these mice were then assessed. Transfection of 253J B-V cells with miR-145 induced apoptosis and suppression of cell migration in vitro. Western blotting showed that the levels of c-Myc, socs7, FSCN1, E-cadherin, β-catenin, and catenin δ-1 were decreased and that the PI3K/Akt and Erk1/2 signaling pathways were increased in compensatory fashion. In vivo, mice treated with miR-145 showed 76% inhibition of tumor growth, with a significant prolongation of animal survival (p = 0.0183 vs. control). Western blotting showed that both apoptosis and cell motility-related genes were significantly decreased as seen in vitro. Furthermore, PI3k/Akt and Erk1/2 signaling pathways, which were activated in a compensatory manner in vitro, were decreased in vivo. Intravesical administration of exogenous miR-145 was thus concluded to be a valid therapy for bladder cancer in this human bladder cancer xenograft model.