摘要
目的探讨膀胱肿瘤导向肽NYZL1的功能及体内外分子动力学特征。方法采用竞争性抑制法检测其与细胞结合,噻唑蓝(MTr)法确定NYZLI的抗肿瘤功能;细胞免疫荧光技术、组织免疫荧光技术和活体荧光成像技术研究NYZL1-异硫氰酸荧光素(FITC)分子探针与肿瘤细胞结合的内化作用及探针在肿瘤微循环和全身器官的动态分布的动力学特征。结果单纯加入不同浓度组NYZL1-FITC荧光值为4/6.00±13.20、966.85±20.20和1751.45±16.40,先加入NYZL1再加入NYZL1-FITC荧光值为180.45±12.40、696.30±16.30、1253.24±11.50,组间差异有统计学意义(P〈0.05);在25μmol/L和100μmol/L浓度下NYZL1和NYZL1-FITC组的吸光度(A)值分别为0.613±0.055、0.646±0.061和0.647±0.063、0.664±0.070,计算各组相对抑制率均低于30%;探针进入细胞,5min出现绿色荧光,10-20min时细胞核显影,4h细胞内荧光强度最强;静脉注射裸鼠后,探针一部分经肝胆循环和肾脏排泄,另外一部分经血液循环向肿瘤组织靶向浓聚,4-6h肿瘤组织浓聚达高峰,但探针在组织中的分布与肿瘤的血管分布密切相关,探针能够同时标记肿瘤血管和细胞。结论NYZL1不具有抗肿瘤效应,能够携带荧光染料进行膀胱肿瘤的靶向光学分子成像。
Objective To study the functions and molecular dynamics characteristics of bladder tumor targeting peptide NYZL1.Methods Methyl thiazol tetrazolium(MTr)method was used to deter-mine the function of NYZL1.Immunofluorescenee techniques,tissue immunofluorescenee techniques and in vivo fluorescence imaging technology were used to research the internalization of NYZL1-fluoreseein Isothiocyanate(FITC)molecular probe in cells,and the distribution and kinetics of molecular probes in the tumor microclrculation and organ in vivo.Results NYZL1-FITC fluorescence separately with differ-ent concentrations is 416.00±13.20,966.85±20.20 and 1751.45±16.40,while fluorescence to join NYZL1 added NYZL1-FITC is 180.45±12.40,696.30±16.30,1253.24±11.50.There are signifi-cant differences between the groups(P〈0.05).In 25μmol/L and 100μmol,/L concentration of NYZL1 and NYZL1-FITC,optical density is0.613±0.055,0.646±0.061 and 0.647±0.063,0.664±0.070 with the relative inhibition rate of less than 30%;The probe into the cell,a green phosphor 5 min,10-20 min the developing nuclei,the fluorescence intensity of the strongest cell on 4 h;After intravenous in-jection of the probe,part of the probe were excreted by the liver-gallbladder and kidney,part of it were targeted uptake to the tumor tissue via the blood circulation,after 4-6 h tumor tissue uptake reached a maximum peak,but the distribution of the probe in tissue related with the tumor blood vessels and mark with the tumor blood vessels and the ceils simultaneously.Conclusion NYZL1 does not have anti-tumor effect,it can carry a fluorescent dye targeting to bladder tumor for optical molecular imaging,will be a goodtargeted carrier in bladder cancer diagnosis and targeted therapy.
作者
苏西西
杨晓峰
张帆
庞建智
闫三华
罗俊茜
罗芳
Su Xixi;Yang Xiaofeng;Zhang Fan;Pang Jianzhi;Yan Sanhua;Luo Junqian;Luo Fang(The First Clinical Medical College,Shanxi Medical University,Taiyuan 030001,China;Departmenut of Urology,the First Hospital Affiliated to Shanxi Medical University,Taiyuan 030001,China;Department of Microbiology and Immunology,Shanxi Medical University,Taiyuan 030001,China)
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2016年第12期2654-2657,共4页
Chinese Journal of Experimental Surgery
基金
国家自然科学基金(81172444)
关键词
膀胱肿瘤
非肌层浸润性肿瘤
肿瘤导向肽
分子动力学
Bladder cancer
Non-muscle invasive tumors
Tumor targeting peptide
Mo-lecular dynamics
