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黄议莹,李喆,潘信利,黄媛林,胡文进,李菲,王巧贞,黄庶识,周晓莹,温文胜.产体外抗鼻咽癌物质红树林土壤细菌筛选及其活性成分分析[J].广西科学,2022,29(5):846-853. [点击复制]
- HUANG Yiying,LI Zhe,PAN Xinli,HUANG Yuanlin,HU Wenjin,LI Fei,WANG Qiaozhen,HUANG Shushi,ZHOU Xiaoying,WEN Wensheng.Screening of Mangrove Soil Bacteria Producing in Vitro Anti-nasopharyngeal Carcinoma Substance and Analysis of Its Active Components[J].Guangxi Sciences,2022,29(5):846-853. [点击复制]
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| 产体外抗鼻咽癌物质红树林土壤细菌筛选及其活性成分分析 |
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黄议莹1, 李喆2, 潘信利2, 黄媛林2, 胡文进3, 李菲2, 王巧贞2, 黄庶识2, 周晓莹4, 温文胜1
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| (1.广西医科大学第一附属医院耳鼻咽喉头颈外科, 广西南宁 530021;2.广西科学院, 广西海洋天然产物与组合生物合成化学重点实验室, 广西南宁 530007;3.广西科学院, 国家非粮生物质能源工程技术研究中心, 非粮生物质酶解国家重点实验室, 广西生物炼制重点实验室, 广西生物质工程技术研究中心, 广西南宁 530007;4.广西医科大学生命科学研究院, 广西南宁 530021) |
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| 摘要: |
| 研究一株通过活性筛选获得的、具有抗鼻咽癌(Nasopharyngeal Carcinoma,NPC)活性物质生产潜力的细菌，对其所产活性物质进行分离纯化，以探讨其抗肿瘤研发价值。以鼻咽癌肿瘤细胞株TW03和5-8F作为测试对象，利用细胞计数试剂盒(Cell Counting Kit-8,CCK8)法从广西北海采集的红树林土壤中筛选出具有体外抗鼻咽癌活性物质生产潜力的细菌，通过基因分析、细菌形态以及生理生化特性对活性菌株进行分类鉴定，通过柱层析分离和活性测试追踪法从菌株的发酵液中获得具有体外抗鼻咽癌活性的化合物，并应用核磁共振波谱和低分辨质谱鉴定其结构。结果表明，经16S rRNA和看家基因(atpD和rpoB)的多位点序列分析鉴定菌株归属于链霉菌，命名为Streptomyces sp.MCCG 218。从该菌株的发酵液中分离得到一个活性化合物 1，经图谱分析和文献比对确认该化合物为巴弗洛霉素A1衍生物(17,18-dehydro-24-demethyl-bafilomycin A1)，其对鼻咽癌细胞株TW03和5-8F表现出较强的细胞增殖抑制活性，半抑制浓度(Half Inhibitory Concentration,IC50)值分别为2.7 μmol/L和9.2 μmol/L。红树林来源链霉菌MCCG 218能够生产具有抑制鼻咽癌细胞增殖活性的化合物巴弗洛霉素A1衍生物。 |
| 关键词: 广西红树林 链霉菌 分离鉴定 抗鼻咽癌 巴弗洛霉素A1衍生物 |
| DOI：10.13656/j.cnki.gxkx.20221116.004 |
| 投稿时间：2021-08-04修订日期：2021-10-12 |
| 基金项目:国家地区科学基金项目(81860479，82260689)，广西海洋天然产物与组合生物合成化学重点实验室运行费项目(17-259-74)，广西科学院基本业务费(2019YBJ101，2018YBJ303)，广西科技计划-科技基地和人才专项(2021AC19380)，广西青年科学基金项目(2018GXNSFBA050021，2018GXNSFBA281028，2021GXNSFBA220020)资助。 |
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| Screening of Mangrove Soil Bacteria Producing in Vitro Anti-nasopharyngeal Carcinoma Substance and Analysis of Its Active Components |
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HUANG Yiying1, LI Zhe2, PAN Xinli2, HUANG Yuanlin2, HU Wenjin3, LI Fei2, WANG Qiaozhen2, HUANG Shushi2, ZHOU Xiaoying4, WEN Wensheng1
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| (1.Department of Otolaryngology-Head and Neck Surgery, the First Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi, 530021, China;2.Guangxi Key Laboratory of Marine Nature Products and Combinatorial Biosynethesis Chemistry, Guangxi Academy of Sciences, Nanning, Guangxi, 530007, China;3.National Engineering Research Center for Non-Food Biorefinery, State Key Laboratory of Non-Food Biomass and Enzyme Technology, Guangxi Key Laboratory of Bio-refinery, Guangxi Biomass Engineering Technology Research Center, Guangxi Academy of Sciences, Nanning, Guangxi, 530007, China;4.Life Science Institute, Guangxi Medical University, Nanning, Guangxi, 530021, China) |
| Abstract: |
| A strain of bacteria with anti-Nasopharyngeal Carcinoma (NPC) active substance production potential obtained by activity screening was studied,and the active substances produced were isolated and purified to explore its anti-tumor research and development value.The nasopharyngeal carcinoma cell lines TW03 and 5-8F were used as the test objects.The Cell Counting Kit-8 (CCK8) method was used to screen the bacteria with the potential to produce anti-nasopharyngeal carcinoma active substances in vitro from the mangrove soil collected from Beihai,Guangxi.The active strain was classified and identified by gene analysis,bacterial morphology and physiological and biochemical characteristics.The compounds with anti-nasopharyngeal carcinoma activity in vitro were obtained from the fermentation broth of strain by column chromatography separation and activity test tracking.The structure was identified by nuclear magnetic resonance spectroscopy and low resolution mass spectrometry.The results showed that the strain was identified to belong to the genus Streptomyces sp.by multilocus sequence analysis of 16 S rRNA and housekeeping genes (atpD and rpoB),named Streptomyces sp.MCCG 218.An active compound 1 was isolated from the fermentation broth of this strain.The compound was identified as a derivative of bafilomycin A1 (17,18-dehydro-24-demethyl-bafilomycin A1) by spectral analysis and literature comparison.It showed strong inhibitory activity of cell proliferation against nasopharyngeal carcinoma cell lines TW03 and 5-8F.The Half Inhibitory Concentration (IC50) value was 2.7 μmol/L and 9.2 μmol/L,respectively.Mangrove-derived Streptomyces sp.MCCG 218 can produce bafilomycin A1 derivatives,which can inhibit the activity on the proliferation of nasopharyngeal carcinoma cells. |
| Key words: Guangxi mangrove Streptomyces isolation and identification anti-nasopharyngeal carcinoma bafilomycin A1 derivative |
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