纳米壳聚糖递送分子信标成像肺癌细胞的研究  

Imaging of lung cancer cells with molecular beacons delivered by chitosan nanoparticles

作　　者：朱海振[1] 俞婕[1] 江飞龙[2] 韩静[3] 李雪涛[3] 陈光朋 李杭[1] 陈正堂[3] 

机构地区：[1]贵州省人民医院肿瘤科,贵阳550002 [2]重庆市中医院,重庆400037 [3]第三军医大学附属新桥医院全军肿瘤研究所,重庆400037

出　　处：《解放军医学杂志》2016年第8期629-635,共7页Medical Journal of Chinese People's Liberation Army

基　　金：国家高技术发展研究计划(“863”计划)(2008AA02Z104);国家自然科学基金面上项目(81071786)~~

摘　　要：目的采用纳米技术和分子信标(MB)技术,研究纳米壳聚糖(CS)作为miR-155 MB载体用于肺癌细胞成像的效果。方法设计并合成锁核酸修饰的miR-155 MB,采用自组装方法合成纳米壳聚糖分子信标复合物(CS-MB)并对其抗DNaseⅠ特性、粒径大小、zeta电位等理化性质进行检测。以CS作为载体转染miR-155 MB,激光共聚焦检测miR-155 MB对肺癌细胞中的miR-155的识别能力并进行成像,采用实时荧光定量PCR(q RT-PCR)进行进一步验证,以随机序列分子信标(RS MB)作为阴性对照。结果按照7:1的质量比合成的CS-MB,其包封率高,能抵抗DNaseⅠ的降解,粒径小,带正电荷,适合基因转染。CS转染miR-155 MB后可在肺癌细胞中看到较强的红色荧光,RS MB组未检测到荧光信号(P<0.05),且荧光信号强弱趋势与实时荧光定量PCR检测的miR-155表达趋势较一致。结论 CS可作为miR-155 MB载体检测肺癌细胞中miR-155的表达并进行肺癌细胞成像,为肺癌的诊断提供了新思路和新技术。Objective To investigate the effect of a method for imaging lung cancer cells using nanotechnology and molecular beacon（MB） that identifies miR-155 and is delivered by chitosan nanoparticles（CS）. Methods The miR-155 MB modified by locked nucleic acids（LNAs） was designed and synthesized. The CS-MB complex was synthesized by self-assembly method and tested for its physicochemical properties including anti-DNase Ⅰ features, particle size, zeta potential and so on. The miR-155 MB was transfected with CS as vectors. The abilities of miR-155 MB to identify miR-155 and thus to image lung cancer cells were determined by confocal microscopy. Furthermore, the miR-155 expression levels were detected by q RT-PCR to validated the effect of miR-155 MB. The random sequence molecular beacon（RS MB） was set as a negative control. Results The CS-MB complex at the weight ratio of 7：1 was best suited for transfection due to its high encapsulation rate, resistance to the degradation by DNase Ⅰ, small particle size and positive charge. Relatively strong red fluorescence could be detected in the lung cancer cells after transfection of miR155 MB, while that could not be detected in the RS MB group（P〈0.05）. Moreover, the changing trend in the fluorescence intensity was consistent with that in the miR-155 expression levels. Conclusion CS nanoparticles can be used as vectors to deliver miR-155 MB for miR-155 identification and lung cancer cell imaging, thus providing new ideas and novel technique for lung cancer diagnosis.

关 键 词：肺肿瘤 纳米壳聚糖 分子信标 微RNA-155 

分 类 号：R734.2[医药卫生—肿瘤]