10-羟基喜树碱的光谱性质、 抗肿瘤活性及应用于细胞标记研究

doi:10.7503/cjcu20140575

高等学校化学学报 ›› 2014, Vol. 35 ›› Issue (11): 2324.doi: 10.7503/cjcu20140575

10-羟基喜树碱的光谱性质、抗肿瘤活性及应用于细胞标记研究

丁国斌¹(), 李彬春¹, 郭轶², 徐力²()

1. 山西大学化学生物学与分子工程教育部重点实验室, 生物技术研究所, 太原 030006
2. 吉林大学分子酶学工程教育部重点实验室, 长春 130012

收稿日期:2014-06-25 出版日期:2014-11-10 发布日期:2014-10-09
作者简介:联系人简介: 丁国斌, 男, 博士, 讲师, 主要从事纳米生物学研究. E-mail:dinggb2012@sxu.edu.cn; 徐力, 女, 博士, 教授, 主要从事酶与纳米生物技术研究. E-mail:xuli@jlu.edu.cn
基金资助:
国家自然科学基金(批准号: 81271697, 30970719)、山西省基础研究项目青年科技研究基金(批准号: 2013021014-1)和山西大学科研启动资金(批准号: 020651901007)资助

Spectral Properties and Antitumor Activity of 10-Hydroxycamptothecin and Its Application in Cell Labeling^†

DING Guobin^1,^*(), LI Binchun¹, GUO Yi², XU Li^2,^*()

1. Institute of Biotechnology, Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education,Shanxi University, Taiyuan 030006, China
2. Key Laboratory for Molecular Enzymolozy and Engineering, Ministry of Education,Jilin University, Changchun 130012, China

Received:2014-06-25 Online:2014-11-10 Published:2014-10-09
Contact: DING Guobin,XU Li E-mail:dinggb2012@sxu.edu.cn;xuli@jlu.edu.cn
Supported by:
† Supported by the National Natural Science Foundation of China(Nos.81271697, 30970719), the Science Foundation for Youths of Shanxi Province, China(No.2013021014-1) and the Scientific Research Starting Foundation of Shanxi University, China(No.020651901007)

摘要/Abstract

摘要：

通过紫外-可见光谱、荧光光谱和红外光谱等方法研究了药物10-羟基喜树碱(HCPT)的光谱性质. 采用溴化噻唑蓝四氮唑(MTT)法测定了HCPT对3种肿瘤细胞(HeLa, MCF-7和HT1080)的抗肿瘤活性, 其IC₅₀值分别为16.37, 16.73和19.24 μg/mL. 测定了HCPT对正常细胞人胚肾细胞系HEK293T的生长抑制活性, 最高抑制率达88.72%, 表明正常细胞比3种肿瘤细胞对药物HCPT更敏感. 以HeLa细胞为模型, 利用Annexin V-FITC细胞凋亡检测试剂盒研究了HCPT的抗肿瘤作用机制, 发现几乎所有细胞均同时被Annexin V-FITC和碘化丙啶(PI)染色, 细胞膜为绿色, 而细胞核为红色, 表明HCPT诱导了HeLa细胞的晚期凋亡. 通过荧光显微镜观察了HCPT在HeLa细胞中的分布, 为其在细胞标记中的应用提供了科学依据.

关键词: 10-羟基喜树碱, 光谱性质, 抗肿瘤活性, 细胞凋亡, 细胞标记

Abstract:

The spectral properties of 10-hydroxycamptothecin(HCPT) were investigated by UV-Vis spectrophotometer, fluorescence spectrophotometer and fourier transform infrared spectroscopy. The antiproliferative activities of HCPT against three tumor cell lines, human cervical can-cer cell line(HeLa), human breast cancer cell line(MCF-7) and human fibrosarcoma cell line(HT1080), were determined using an MTT assay, and the IC₅₀ values of HCPT to the three tumor cell lines were 16.37, 16.73 and 19.24 μg/mL, respectively. The growth inhibitory effect of HCPT on a normal cell line, HEK293T(human embryonic kidney cells 293T), was also investigated, the highest inhibitory rate reached 88.72% at an HCPT concentration of 40 μg/mL, indicating that the normal cell line HEK293T is more sensitive to HCPT in comparison to the three tumor cell lines. Using HeLa cell line as a model, Annexin V-FITC apoptosis detection kit was employed to demonstrate the antitumor mechanism of HCPT. Almost all the cells were simultaneously stained by Annexin V-FITC and PI, the cell membrane was green while the nucleus was red, which indicated that HCPT induced the late apoptosis process of HeLa cells. The distribution of HCPT in HeLa cells was observed by fluorescence microscopy, which provides scientific basis for its application in cell labeling.

Key words: 10-Hydroxycamptothecin, Spectral property, Antitumor activity, Apoptosis, Cell labeling

中图分类号:

O629.3

TrendMD:

丁国斌, 李彬春, 郭轶, 徐力. 10-羟基喜树碱的光谱性质、抗肿瘤活性及应用于细胞标记研究. 高等学校化学学报, 2014, 35(11): 2324.

DING Guobin, LI Binchun, GUO Yi, XU Li. Spectral Properties and Antitumor Activity of 10-Hydroxycamptothecin and Its Application in Cell Labeling^†. Chem. J. Chinese Universities, 2014, 35(11): 2324.

图/表 5

Fig.1 UV-Vis spectra of HCPT solutions with different concentrations c/(μg·mL-1): a—g. 1, 5, 10, 20, 30, 40, 50.

Fig.2 Fluorescence spectra of HCPT solutions with different concentrations Inset: the images of HCPT solutions(1, 20 and 50 μg/mL) under an ultraviolet lamp(λex=365 nm).

Fig.3 In vitro antitumor activities of HCPT solutions (A) HCPT solutions with different concentrations against HeLa, MCF-7 and HT1080 cells; (B) growth inhibitory effect of HCPT on HEK293T cells after 48 h incubation at 37 ℃, values are averages of three replicates±SD(standard deviation).

Fig.4 Fluorescence microscopy images of HeLa cells treated with control cell medium(A) and HCPT(40 μg/mL)(B) for 24 h followed by staining with an annexin V-FITC apoptosis detection kit

Fig.5 Fluorescence microscopy images of HeLa cells incubated with control cell medium(A) and HCPT(40 μg/mL)(B) for 12 h

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10-羟基喜树碱的光谱性质、抗肿瘤活性及应用于细胞标记研究

Spectral Properties and Antitumor Activity of 10-Hydroxycamptothecin and Its Application in Cell Labeling^†

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10-羟基喜树碱的光谱性质、 抗肿瘤活性及应用于细胞标记研究

Spectral Properties and Antitumor Activity of 10-Hydroxycamptothecin and Its Application in Cell Labeling†

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10-羟基喜树碱的光谱性质、抗肿瘤活性及应用于细胞标记研究

Spectral Properties and Antitumor Activity of 10-Hydroxycamptothecin and Its Application in Cell Labeling^†