地皮菜中一种新型水应激蛋白基因的克隆表达及其抗人结肠癌细胞SW480的作用

doi:10.7506/spkx1002-6630-201413029

食品科学

地皮菜中一种新型水应激蛋白基因的克隆表达及其抗人结肠癌细胞SW480的作用

郭松佳，单树花，晋小婷，李宗伟，宋莉，李卓玉

1.化学生物学与分子工程教育部重点实验室，山西大学生物技术研究所，山西太原 030006；2.山西医科大学汾阳学院
医学检验系，山西汾阳 032200；3.太原师范学院生物系，山西太原 030031

出版日期:2014-07-15 发布日期:2014-07-18

Cloning and Expression of a Novel Water Stress Protein from Nostoc commune Vauch. and Its Inhibitory Effect on the Proliferation of Human Colon Cancer SW480 Cells

GUO Song-jia, SHAN Shu-hua, JIN Xiao-ting, LI Zong-wei, SONG Li, LI Zhuo-yu

1. Laboratory for Chemical Biology and Molecular Engineering, Ministry of Education, Institute of Biotechnology, Shanxi University,
Taiyuan 030006, China; 2. Department of Medical Laboratory, Fenyang College of Shanxi Medical University, Fenyang 032200,
China; 3. Department of Biology, Taiyuan Normal University, Taiyuan 030031, China

Online:2014-07-15 Published:2014-07-18

摘要/Abstract

摘要：

利用聚合酶链式反应（polymerase chain reaction，PCR）扩增出地皮菜（Nostoc commune Vauch.）一种新型水应激蛋白（water stress protein，WSP）的基因（GenBank 序列号：KF003026），连接到原核表达载体pET28a上，测序鉴定后转化入大肠杆菌BL21，获得一种新型水应激蛋白表达工程菌；经异丙基-β-D-硫代半乳糖苷（isopropyl-β-D-thiogalactopyranoside，IPTG）诱导表达及镍离子亲和层析柱（Ni-NTA）纯化后，得到了带6×His标签、分子质量大小为40 kD的可溶性目的蛋白，称其为重组水应激蛋白1（recombinant water stress protein 1，Re-WSP1）。细胞增殖抑制实验结果表明，Re-WSP1蛋白可以抑制结肠癌细胞SW480的增殖，而对正常结肠上皮FHC细胞无明显作用；DAPI细胞核染色结果表明，该蛋白能够诱导SW480细胞凋亡小体的形成；Western blotting结果显示，Re-WSP1蛋白能够促进Procaspase-3和Procaspase-8的活化剪切。以上结果表明：Re-WSP1蛋白能够明显的抑制结肠癌SW480细胞的增殖，并可能通过Caspase依赖途径诱导SW480细胞凋亡。

关键词: 地皮菜, 重组水应激蛋白, 表达与纯化, 结肠癌, 细胞凋亡

Abstract:

Water stress protein (WSP) gene was amplified by polymerase chain reaction (PCR) from the Nostoc commoune
Vauch. genome (Genbank NO. KF003026), and cloned into the prokaryotic expression vector pET28a. After being
sequenced, the recombinant plasmid pET28a-WSP was transformed into E.coli BL21, and recombinant water stress protein 1
(Re-WSP1) was induced to express by isopropyl-β-D-thiogalactoside (IPTG). The soluble 6 × histidine (His)-tagged fusion
Re-WSP1 was further purified by Ni+-NTA affinity chromatography. The molecular weight of the fused Re-WPS1 was 40 kD
as expected. When applied to cells in culture, Re-WSP1 could significantly suppress the growth of SW480 colon cancer
cells, but had a negligible effect on that of FHC normal colon epithelial cells. In addition, Re-WSP1-treated SW480 cells
displayed the apoptotic characteristics of chromatin condensation and typical fragmented nuclei, as demonstrated by DAPI
staining. Re-WSP1 could also activate procaspase-8 and procaspase-3, resulting in their cleavage into active fragments.
Taken together, Re-WSP1 has shown obvious anti-colon cancer effects on SW480 cells, and induced apoptosis by caspasedependent
pathway in SW480 cells.

Key words: Nostoc commune Vauch., recombinant water stress protein 1 (Re-WSP1), expression and purification, colon cancer, cell apoptosis

郭松佳，单树花，晋小婷，李宗伟，宋莉，李卓玉. 地皮菜中一种新型水应激蛋白基因的克隆表达及其抗人结肠癌细胞SW480的作用[J]. 食品科学, doi: 10.7506/spkx1002-6630-201413029.

GUO Song-jia, SHAN Shu-hua, JIN Xiao-ting, LI Zong-wei, SONG Li, LI Zhuo-yu. Cloning and Expression of a Novel Water Stress Protein from Nostoc commune Vauch. and Its Inhibitory Effect on the Proliferation of Human Colon Cancer SW480 Cells[J]. FOOD SCIENCE, doi: 10.7506/spkx1002-6630-201413029.

[1]	薛宏坤，谭佳琪，李倩，唐劲天. 原花青素B2诱导乳腺癌MCF-7细胞凋亡及其作用机制[J]. 食品科学, 2021, 42(9): 91-99.
[2]	杨成林，黄超，刘娟，黄卫梅，袁志航，易金娥，梁曾恩妮，邬静. 桦木酸对T-2毒素诱导小鼠肝损伤的保护作用[J]. 食品科学, 2021, 42(9): 107-114.
[3]	李宁宁，李钊，赵圣明，马汉军，张文振，康壮丽，朱明明，王正荣，何鸿举. 地皮菜添加量对鸡胸肉糜凝胶特性的影响[J]. 食品科学, 2021, 42(2): 53-59.
[4]	何萍萍，郑雅君，郑明静，姜泽东，杜希萍，朱艳冰，倪辉，李清彪. 红毛藻多糖对H2O2诱导的Caco-2细胞氧化损伤的保护作用[J]. 食品科学, 2021, 42(17): 113-120.
[5]	王新怡，张一敏，梁荣蓉，董鹏程，罗欣，毛衍伟. 过氧化物氧化还原酶6调控肉品嫩度的研究进展[J]. 食品科学, 2021, 42(11): 236-243.
[6]	杜月梅，韩何丹，郭卓雨，高丽萍. 葡萄籽原花青素对顺铂诱导小鼠睾丸支持细胞TM4细胞凋亡的影响[J]. 食品科学, 2020, 41(9): 98-104.
[7]	郝帅，李爽，王静，吴婷婷，张佳雯，王成涛. 基于转录组技术研究藻蓝蛋白在抑制非小细胞肺癌中的调控作用[J]. 食品科学, 2020, 41(5): 93-101.
[8]	孙金龙，师希雄，黄峰，韩玲，陈骋，岳建伟. 藏羊肉宰后成熟过程中热休克蛋白27对肌原纤维蛋白及细胞凋亡酶的影响[J]. 食品科学, 2020, 41(3): 24-29.
[9]	田艳，邓放明，赵玲艳，廖安，赖灯妮，陈秋佳. 高菜硫代葡萄糖苷提取物对人结肠癌细胞HCT116的抑制作用[J]. 食品科学, 2020, 41(23): 172-180.
[10]	林杨, 杨平, 张琦, 郎宇曦, 邓皓天, 孟宪军. 蓝莓花色苷对脂多糖诱导RAW 264.7细胞炎症及结肠癌细胞增殖、凋亡的影响[J]. 食品科学, 2020, 41(13): 133-140.
[11]	侯颖，马文瑞，程雅文，郑浩然，谭之磊，贾士儒. 聚赖氨酸偶联活性氧诱导酿酒酵母细胞凋亡[J]. 食品科学, 2020, 41(10): 81-87.
[12]	郭方明，李述刚，宋关玲，丁玉松，冯刚玲，牛强，徐上知，胡云华. 葡萄籽提取物原花青素诱导人食管癌细胞ECA109凋亡[J]. 食品科学, 2019, 40(9): 115-121.
[13]	袁锦莹，孙万成，罗毅皓，丁建霞. 牦牛酥油支链脂肪酸对人乳腺癌细胞抑制的转录组学分析[J]. 食品科学, 2019, 40(9): 195-200.
[14]	孙思明，徐宏伟，郭凯元，郭梦冉，崔胜男，崔同，檀建新. 山楂原花青素对人肝癌SMMC-7721细胞增殖和凋亡的影响[J]. 食品科学, 2019, 40(5): 189-196.
[15]	张立超，杨宇菲，李汉卿，剌晓琴，李卓玉,. 胡麻粕多酚对肠癌细胞凋亡的促进作用[J]. 食品科学, 2019, 40(23): 176-182.

地皮菜中一种新型水应激蛋白基因的克隆表达及其抗人结肠癌细胞SW480的作用

Cloning and Expression of a Novel Water Stress Protein from Nostoc commune Vauch. and Its Inhibitory Effect on the Proliferation of Human Colon Cancer SW480 Cells

RichHTML

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价