奥曲肽修饰的壳聚糖纳米递送分子信标成像肺癌研究  

Imaging of lung cancer with molecular beacons delivered by octreotide-modified chitosan nanoparticles

作　　者：马雪 吴婧 张红丽[2] 李勇[2] 宋娟 李源丽 鲁亮[2] 朱海振[2] MA Xue;WU Jing;ZHANG Hongli;LI Yong;SONG Juan;LI Yuanli;LU Liang;ZHU Haizhen(Graduate Institute of Guizhou University of Chinese Medicine,Guiyang 550000,China;Department of Oncology,Guizhou Provincial People's Hospital,NHC Key Laboratory of Pulmonary Immunological Diseases of Guizhou Provincial People's Hospital,Guizhou Cancer Center)

机构地区：[1]贵州中医药大学研究生院,550000 [2]贵州省人民医院肿瘤科,贵州省人民医院肺脏免疫性疾病诊治国家重点实验室,贵州省癌症中心

出　　处：《天津医药》2024年第1期61-67,共7页Tianjin Medical Journal

基　　金：国家自然科学基金地区基金项目(82260357);贵州省科技计划项目(黔科合基础[2020]1Y428,黔科合支撑[2021]一般093)。

摘　　要：目的 探究通过奥曲肽(OCT)修饰的壳聚糖(CS)miR-155分子信标(miR-155-MB)纳米(CS-miR-155-MB-OCT)识别miR-155并成像肺癌细胞用于肺癌早期诊断。方法 通过尾静脉注射A549肺癌细胞建立肺移植瘤裸鼠模型;通过鼻腔滴入Cre腺病毒,分别在滴入腺病毒后的4、6、8、12周处死小鼠建立LSL K-ras G12D转基因小鼠肺部的非典型增生、腺瘤、原位癌、肺腺癌的不同病变时期肺腺癌模型;取肺组织行HE染色观察。免疫组化染色检测肺移植瘤组织及不同病变时期转基因小鼠肺组织生长抑素受体2(SSTR2)的表达;实时荧光定量PCR检测不同病变时期转基因小鼠肺组织miR-155的表达。在肺移植瘤裸鼠模型中,尾静脉注射CS-miR-155-MB或CS-miR-155-MB-OCT;转基因小鼠模型中,尾静脉注射CS-miR-155-MB-OCT;活体成像仪检测肺移植瘤裸鼠及转基因小鼠不同病变时期肺部荧光信号;制作肺组织冰冻切片,激光共聚焦扫描显微镜(CLSM)检测荧光信号来源。结果 HE染色显示成功构建了肺移植瘤裸鼠模型及转基因小鼠肺部的非典型增生、腺瘤、原位癌、肺腺癌的不同病变时期肺癌模型。肺移植瘤及不同病变时期病变组织均表达SSTR2。转基因小鼠模型中,随着疾病进展,miR-155表达逐渐升高(P<0.05)。在肺移植瘤裸鼠模型中,CS-miR-155-MB-OCT组肺部荧光信号强于CS-miR-155-MB组(P<0.05);转基因小鼠模型中,随着肺癌的进展,荧光信号逐渐增强(P<0.05);将肺组织再次成像后发现荧光信号来自肺部,CLSM发现荧光信号来自肿瘤细胞及部分正常肺泡上皮细胞。结论 CS-miR-155-MB-OCT能够根据产生的荧光强度变化动态反映肺癌的发生发展,从而为肺癌的早期诊断提供新技术。Objective To investigate the identification of octreotide(OCT)modified chitosan(CS)miR-155 molecular beacon nanoparticles(CS-miR-155-MB-OCT)and imaging of lung cancer cells for the early screening of lung cancer.Methods A nude mouse model of lung transplantation tumor was established by injecting A549 lung cancer cells into tail veins to establish lung xenograft models.Cre adenovirus was injected through nasal cavity,and mice were killed at 4,6,8 and 12 weeks after adenovirus injection to establish lung cancer models of atypical hyperplasia,adenoma,carcinoma in situ and adenocarcinoma of lung in LSL K-ras G12D transgenic mice at different pathological stages.Lung tissue samples were taken and observed by HE staining.Immunohistochemistry were used to detect the expression of somatostatin receptor 2(SSTR2).Real-time fluorescence quantitative PCR was used to detect miR-155 expression levels in lung xenograft models and transgenic mice at different stages of lung cancer.Then CS-miR-155-MB and CS-miR-155-MB-OCT were injected via tail vein in lung xenograft models.CS-miR-155-MB-OCT was injected via tail vein in transgenic mice models.The fluorescence signals of lung in nude mice and transgenic mice at different disease stages were imaged by living imaging system.Frozen slices of lung tissue were made.The source of fluorescence signal was detected by laser confocal scanning microscope(CLSM).Results HE staining showed that lung transplantation tumor models and lung cancer models of atypical hyperplasia,adenoma,carcinoma in situ and lung adenocarcinoma at different pathological stages were successfully constructed.Immunohistochemical analysis showed somatostatin receptor 2(SSTR2)was expressed in transplanted lung tumor and tissue at different pathological stages.In transgenic mouse models,the expression of miR-155 was gradually increased as the disease progressed(P<0.05).In lung xenograft models,the fluorescence signals were significantly higher in the CS-miR-155-MB-OCT group than those of the CS-miR-155-MB group(P<0.05).

关 键 词：壳聚糖 奥曲肽 肺肿瘤 分子信标 MIR-155 分子成像 

分 类 号：R734.2[医药卫生—肿瘤]