荧光素酶生物发光报告体系的优化及在肝癌荷瘤裸鼠模型中的应用*

doi:10.3969/j.issn.1672-5069.2019.04.008

实用肝脏病杂志 ›› 2019, Vol. 22 ›› Issue (4): 482-485.doi: 10.3969/j.issn.1672-5069.2019.04.008

荧光素酶生物发光报告体系的优化及在肝癌荷瘤裸鼠模型中的应用^*

王爽, 许坚吉, 刘晓霓, 陈德喜

100069北京市首都医科大学附属北京地坛医院检验科（王爽）;
首都医科大学附属北京佑安医院北京市肝病研究所（许坚吉,刘晓霓,陈德喜）

收稿日期:2018-09-25 出版日期:2019-07-10 发布日期:2019-07-19
通讯作者: 刘晓霓：E-mail：lxnlxm@126.com
作者简介:王爽,女,33岁,医学博士,助理研究员。主要从事肝癌分子机制研究。E-mail：wsh1029@126.com
基金资助:
*国家自然科学基金资助项目（81672026）; 北京市卫生系统高层次卫生技术人才培养资助项目（2015-3-101）; 首都卫生发展科研专项项目（2018-1-1151）; 北京市自然科学基金资助项目（7192084）

Optimization of luciferase bioluminescence reporting system and its application in xenografted human HepG2 cell transplantation in nude mice

Wang Shuang, Xu Jianji, Liu Xiaoni, et al

Clinical Laboratory,Beijing Ditan Hospital,Capital Medical University,Beijing 100069

Received:2018-09-25 Online:2019-07-10 Published:2019-07-19

摘要/Abstract

摘要： 目的优化荧光素酶生物发光报告体系条件,提高基于荧光素酶生物发光活体成像技术在活体肿瘤研究中的检测灵敏性和应用潜力。方法在一定数目稳定表达荧光素酶的肝癌细胞（HepG2-Luc）基础上（固定荧光素酶量）进行体外luciferin和ATP的最佳浓度优化,以此优化条件进行荷瘤裸鼠活体成像灵敏度检测和发光强度分析。结果 Luciferin的最佳浓度是1.25 mg/ml,ATP缓冲液的最佳浓度是12.5 μM。活体成像实验显示,未使用优化体系时,将2×10⁶肝癌细胞皮下注射后检测到的光强度为484645,使用优化体系进行检测的发光强度为1483024;对荷瘤裸鼠进行检测时,未使用优化体系时的实验鼠与对照鼠的信号强度比值为0.11,而优化后的比值增加为0.67,说明以此配比优化的荧光素酶报告系统能够明显提高裸鼠体内肿瘤的检出灵敏度,延长发光时间。结论应用我们建立的荧光素酶生物发光报告体系优化方法可以显著提高肝癌荷瘤裸鼠活体成像检测的灵敏度。

关键词: HepG2细胞, 活体成像技术, 生物发光, 荧光素酶, 裸鼠

Abstract: Objective To optimize the luciferase bioluminescence reporting system and to investigate its application in xenografted human HepG2 cell transplantation in nude mice.Methods A certain number of HepG2-Luc cells with stable expression of luciferase（fixed luciferase amount） were conducted for optimization of luciferin and ATP concentration. The sensitivity and luminescence intensity of the optimized substrate and ATP concentration were analyzed in vivo for the tumor cell imaging in nude mice.Results The optimal concentration of luciferin was 1.25 mg/ml and the optimal concentration of ATP buffer was 12.5 μM in in vivo imaging experiments;the intensity of subcutaneous injection of 2×10⁶ HepG2-Luc cells was 484645,while it became 1483024 when the optimized system was used;the ratio of signal intensity between experimental and control mice was 0.11,while it became 0.67 after the optimized system was used,suggesting that in vivo imaging with this optimized luciferase reporting system could significantly improve the sensitivity of tumor detection and prolong the luminescence time in nude mice. Conclusion An optimized luciferase bioluminescence reporting system is established,which might be used in in vivo study for tumor detections.

Key words: HepG2 cells, In vivo imaging, Bioluminescence, Luciferase, Nude mice

王爽, 许坚吉, 刘晓霓, 陈德喜. 荧光素酶生物发光报告体系的优化及在肝癌荷瘤裸鼠模型中的应用^*[J]. 实用肝脏病杂志, 2019, 22(4): 482-485.

Wang Shuang, Xu Jianji, Liu Xiaoni, et al. Optimization of luciferase bioluminescence reporting system and its application in xenografted human HepG2 cell transplantation in nude mice[J]. JOURNAL OF PRACTICAL HEPATOLOGY, 2019, 22(4): 482-485.

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荧光素酶生物发光报告体系的优化及在肝癌荷瘤裸鼠模型中的应用^*

Optimization of luciferase bioluminescence reporting system and its application in xenografted human HepG2 cell transplantation in nude mice

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