Evaluation of antitumoreffects following tumor necrosis factor-alphagene delivery using nanobubbles and ultrasound
Evaluation of antitumoreffects following tumor necrosis factor-alphagene delivery using nanobubbles and ultrasound
复制标题
使用纳米气泡和超声评估肿瘤坏死因子-α基因递送后的抗肿瘤效果
DOI：
10.1111/j.1349-7006.2011.02084.x
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发表时间：
2011
期刊：
影响因子：
5.7
通讯作者：
Shiro Mori(他4名)
中科院分区：
文献类型：
--
作者：
Satoh Y;Yajima E;Murakami T;Shiro Mori(他4名)
采用纳米泡（NBs）和超声（US）基因传递系统将肿瘤坏死因子（TNF - α）质粒DNA转染到实体小鼠肿瘤中，观察肿瘤坏死因子（TNF - α）的抗肿瘤作用。将表达荧光素酶的小鼠乳腺癌（EMT6）细胞（1 × 106个细胞）于第0天皮下注射到6-7周龄雄性SCID小鼠的侧翼。瘤内注射10微升TNF - α (5 μg/μL)或TNF - α模拟质粒DNA (5 μg/μL)，加/不加NBs （15 μL）和生理盐水，总容积为30 μL，于第2、4、7、9天对肿瘤进行US暴露（频率1 MHz，强度3.0 W/cm2，占空比20%，脉冲数200次，暴露时间60 s）。用活体荧光成像系统和机械卡尺测量肿瘤大小的变化。使用Sonazoid、高频超声成像系统（40 MHz）和免疫组织化学（CD31）进行对比增强超声成像，量化肿瘤血管面积的变化。在mRNA水平上，使用实时定量RT - PCR定量检测TNF - α、caspase - 3和p53的表达。在蛋白水平上，免疫组织化学证实caspase‐3和p53的表达。我们发现，使用NBs和US重复传递TNF - α基因可以导致TNF - α的局部产生。这导致了抗肿瘤作用，包括激活p53依赖的细胞凋亡，降低肿瘤血管密度，抑制肿瘤大小。在这项研究中，我们证明了使用NBs和US将TNF‐α基因递送到肿瘤细胞中的有效性。（癌症科学2011；102:2082-2089）
The antitumor effects of tumor necrosis factor (TNF‐α) were evaluated following transfection of TNF‐α plasmid DNA into solid mouse tumors using the nanobubbles (NBs) and ultrasound (US) gene delivery system. Murine breast carcinoma (EMT6) cells expressing luciferase (1 × 106cells) were injected intradermally into the flanks of 6–7‐week‐old male SCID mice on day 0. Ten microliters of TNF‐α (5 μg/μL) or TNF‐α mock plasmid DNA (5 μg/μL) with/without NBs (15 μL) and saline was injected intratumorally in a total volume of 30 μL, and tumors were exposed to US (frequency, 1 MHz; intensity, 3.0 W/cm2; duty cycle, 20%; number of pulses, 200; and exposure time, 60 s) on days 2, 4, 7, and 9. Changes in tumor size were measured with anin vivobioluminescent imaging system and a mechanical caliper. Changes in tumor vessel area were quantified using contrast‐enhanced US imaging with Sonazoid and a high frequency US imaging system (40 MHz) and immunohistochemistry (CD31). At the mRNA level, expression of TNF‐α, caspase‐3, and p53 were quantified using real‐time quantitative RT‐PCR. At the protein level, expression of caspase‐3 and p53 were confirmed by immunohistochemistry. We show that repeated TNF‐α gene delivery using NBs and US can lead to the local production of TNF‐α. This results in antitumor effects, including activation of p53‐dependent apoptosis, decrease in tumor vessel density, and suppression of tumor size. In this study, we showed the effectiveness of using NBs and US for TNF‐α gene delivery into tumor cells. (Cancer Sci2011; 102: 2082–2089)