| IL-6/STAT3报告基因系统的构建及抑制IL-6/STAT3信号通路的中药单体的筛选及验证 |
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| 引用本文： | 张露露,李晶哲,马琰岩,尚腾飞,王子依,刘长振.IL-6/STAT3报告基因系统的构建及抑制IL-6/STAT3信号通路的中药单体的筛选及验证[J].生物工程学报,2020,36(2):353-361. |
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| 作者姓名： | 张露露 李晶哲 马琰岩 尚腾飞 王子依 刘长振 |
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| 作者单位： | 中国中医科学院医学实验中心 中医药防治重大疾病北京市重点实验室，北京 100700,中国中医科学院医学实验中心 中医药防治重大疾病北京市重点实验室，北京 100700,中国中医科学院医学实验中心 中医药防治重大疾病北京市重点实验室，北京 100700,中国中医科学院医学实验中心 中医药防治重大疾病北京市重点实验室，北京 100700,中国中医科学院医学实验中心 中医药防治重大疾病北京市重点实验室，北京 100700,中国中医科学院医学实验中心 中医药防治重大疾病北京市重点实验室，北京 100700 |
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| 基金项目： | 国家自然科学基金 (Nos. 31170829, 81171762, 81550017)，北京自然科学基金 (No. 7172150)，中国医科院研究项目 (Nos. zz2015015, zz2018006) 资助。 |
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| 摘 要： | 寻找可抑制IL-6/STAT3信号通路的活化从而抑制肿瘤的生长和恶化的中药单体化合物具有重要意义及发展前景。文中通过基因重组技术构建出一种含有STAT3增强子序列和NanoLuc(NLuc)报告基因序列的新表达载体,并进一步建立受STAT3调控并稳定表达NLuc荧光素酶的细胞系,利用该细胞系定量检测多种中药单体化合物对IL-6/STAT3信号通路的调控作用,并对抑制IL-6/STAT3信号通路的中药单体的效果进行验证。酶切鉴定及测序结果表明报告基因表达载体pQCXIP-STAT3-NLuc构建成功。STAT3转录因子的刺激物白细胞介素-6(IL-6)作用于所构建的稳定表达NLuc的细胞系后出现特异性荧光素酶反应,且作用效果呈良好的剂量依赖性,表明受STAT3调控稳定表达NLuc荧光素酶的细胞系构建成功。Western blotting及Real-time PCR实验结果表明所筛选的中药单体化合物石斛碱及粉防己碱可抑制IL-6/STAT3信号通路并显著下调其下游基因Bcl-2及Bcl-x的表达,且作用呈剂量依赖性。综上所述,文中构建了可高效检测STAT3转录活性的报告基因系统,并利用该系统成功地筛选出可抑制IL-6/STAT3信号通路的中药单体化合物,具有一定的理论和应用价值。
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| 关 键 词： | IL-6/STAT3信号系统 NLuc荧光素酶 石斛碱 粉防己碱 |
| 收稿时间： | 2019/9/16 0:00:00 |
Construction of IL-6/STAT3 reporter gene system for screening and validation of traditional Chinese medicine monomers inhibiting IL-6/STAT3 signaling pathway |
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| Lulu Zhang,Jingzhe Li,Yanyan M,Tengfei Shang,Ziyi Wang and Changzhen Liu.Construction of IL-6/STAT3 reporter gene system for screening and validation of traditional Chinese medicine monomers inhibiting IL-6/STAT3 signaling pathway[J].Chinese Journal of Biotechnology,2020,36(2):353-361. |
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| Authors: | Lulu Zhang Jingzhe Li Yanyan M Tengfei Shang Ziyi Wang and Changzhen Liu |
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| Institution: | Beijing Key Laboratory of Research of Chinese Medicine on Prevention and Treatment for Major Diseases, Experimental Research Center, China Academy of Chinese Medical Sciences, Beijing 100700, China,Beijing Key Laboratory of Research of Chinese Medicine on Prevention and Treatment for Major Diseases, Experimental Research Center, China Academy of Chinese Medical Sciences, Beijing 100700, China,Beijing Key Laboratory of Research of Chinese Medicine on Prevention and Treatment for Major Diseases, Experimental Research Center, China Academy of Chinese Medical Sciences, Beijing 100700, China,Beijing Key Laboratory of Research of Chinese Medicine on Prevention and Treatment for Major Diseases, Experimental Research Center, China Academy of Chinese Medical Sciences, Beijing 100700, China,Beijing Key Laboratory of Research of Chinese Medicine on Prevention and Treatment for Major Diseases, Experimental Research Center, China Academy of Chinese Medical Sciences, Beijing 100700, China and Beijing Key Laboratory of Research of Chinese Medicine on Prevention and Treatment for Major Diseases, Experimental Research Center, China Academy of Chinese Medical Sciences, Beijing 100700, China |
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| Abstract: | |
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| Keywords: | IL-6/STAT3 signaling system NLuc luciferase dendrobiine tetrandrine |
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