Paclitaxel tumor-priming enhances siRNA delivery and transfection in 3-dimensional tumor cultures.
Paclitaxel tumor-priming enhances siRNA delivery and transfection in 3-dimensional tumor cultures.
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DOI：
10.1021/mp1004383
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发表时间：
2011-06-06
影响因子：
4.9
通讯作者：
Au JL
中科院分区：
文献类型：
--
作者：
Wong HL;Shen Z;Lu Z;Wientjes MG;Au JL
siRNA癌症治疗剂的临床开发受到实体瘤中较差的间质转运和低效转染的限制。我们已经表明，紫杉醇预处理，通过诱导细胞凋亡，导致扩大的间隙空间，从而提高纳米粒子的交付和运输在肿瘤结节（称为紫杉醇肿瘤引发）和疗效的纳米药物在荷瘤动物。本研究评估紫杉醇肿瘤引发是否改善人口咽癌FaDu细胞的二维和三维培养物中siRNA的递送和转染。我们使用荧光siGLO和共聚焦显微镜来监测转运，并使用生存素siRNA和免疫染色和免疫印迹来监测转染。Survivin是一种化疗诱导的耐药基因/蛋白。将siRNA装载在阳离子脂质体中。结果显示，用50-200 nM紫杉醇预处理（siRNA前24或48小时）增强了siGLO进入单层的总摄取（~ 15%，p<0.05），以及进入三维球状体和肿瘤碎片组织培养物的渗透深度（渗透-深度曲线下的面积大2.1- 2.5倍）。在单层细胞和组织培养中，紫杉醇预处理诱导survivin上调（p<0.05）。单独的Survivin siRNA以剂量依赖性方式降低Survivin水平，并且在紫杉醇预处理后应用Survivin siRNA完全消除紫杉醇诱导的Survivin增加。这些发现表明紫杉醇肿瘤引发不会损害SiRNA的功能。综上所述，紫杉醇肿瘤引发改善了siRNA在肿瘤中的渗透、转染和功能，从而为开发化学siRNA癌症基因治疗提供了有希望和实用的手段。
The clinical development of siRNA cancer therapeutics is limited by the poor interstitial transport and inefficient transfection in solid tumors. We have shown that paclitaxel pretreatment, by inducing apoptosis, causes expansion of the interstitial space and thereby improves nanoparticle delivery and transport in tumor interstitium (referred to as paclitaxel tumor priming) and efficacy of nanomedicines in tumor-bearing animals. The present study evaluated whether paclitaxel tumor priming improves the delivery and transfection of siRNA in 2- and 3-dimensional cultures of human oropharyngeal carcinoma FaDu cells. We used the fluorescent siGLO and confocal microcopy to monitor transport, and used survivin siRNA and immunostaining and immunoblotting to monitor transfection. Survivin is a chemoresistance gene/protein, inducible by chemotherapy. siRNA was loaded in cationic liposomes. The results showed that pretreatment with 50-200 nM paclitaxel (24 or 48 hr before siRNA) enhanced the total uptake of siGLO into monolayers (~15%, p<0.05), and the depth of penetration into 3-dimensional spheroids and tumor fragment histocultures (2.1- to 2.5-times greater area under the penetration-depth curve). In both monolayer cells and histocultures, paclitaxel pretreatment induced survivin upregulation (p<0.05). Survivin siRNA alone decreased the survivin levels in a dose-dependent manner and applying survivin siRNA after paclitaxel pretreatment completely abolished the paclitaxel-induced survivin increases. These findings indicate that paclitaxel tumor priming did not compromise the siRNA functionality. In summary, paclitaxel tumor priming improved the penetration, transfection and functionality of siRNA in tumors, thus offering a promising and practical means to develop chemo-siRNA cancer gene therapy.
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影响因子：
64.8
作者：
Mello, CC;Conte, D
通讯作者：
Conte, D
影响因子：
4.8
作者：
Ling, X;Bernacki, RJ;Li, FZ
通讯作者：
Li, FZ
影响因子：
5.1
作者：
Gurzov, EN;Izquierdo, M
通讯作者：
Izquierdo, M
影响因子：
5.1
作者：
Hasegawa, S;Hirashima, N;Nakanishi, M
通讯作者：
Nakanishi, M
影响因子：
5.7
作者：
Duan, Zhenfeng;Weinstein, Edward J.;Ji, Diana;Ames, Rachel Y.;Choy, Edwin;Mankin, Henry;Hornicek, Francis J.
通讯作者：
Hornicek, Francis J.