| 摘要: |
| 目的：本文旨在研究耐热直接溶血毒素(TDH)在体内和体外对小鼠黑色素瘤细胞B16的抑制作用。方法：通过MTT法、克隆形成试验、凋亡试验、Caspase-8和Caspase-3的活性试验、线粒体膜电位的检测以及体内C57BL/6小鼠荷瘤实验,比较TDH作用于不同细胞的半抑制浓度IC50,评价TDH对小鼠黑色素瘤细胞B16的体内外抑制作用。结果：人结肠上皮细胞NCM460、人肝成纤维细胞LO2、人肝癌细胞SMMC-7721和小鼠黑色素瘤细胞B16在TDH处理24h之后,细胞的半抑制浓度IC50分别为151μg/mL、118μg/mL、54μg/mL和48μg/mL,正常细胞的IC50高出癌细胞近2-3倍。当浓度低于20μg/mL时,TDH以剂量依赖性的方式抑制B16细胞的克隆形成,6mg/kg TDH在移植瘤模型中显著抑制体内肿瘤的生长(P<0.05)。流式细胞术和荧光试剂盒检测表明：20μg/mL的TDH能诱导19.4%的B16细胞发生早期凋亡,并激活Caspase-8和Caspase-3,但不影响线粒体膜电位。结论：TDH具有体内外的抗肿瘤活性,可能通过细胞表面的死亡受体介导的凋亡信号通路引起凋亡,从而发挥抗肿瘤作用。 |
| 关键词: 海洋毒素 耐热直接溶血毒素 抗肿瘤 死亡受体 凋亡 |
| DOI： |
| 分类号:R979.1 |
| 基金项目: |
|
| The antitumor effects of Vibrio parahaemolyticus thermostable direct hemolytic toxin on melanoma |
|
YANG Lan-zhu1, LI Shi-yi2, ZHENG Wen-jing2, LI Jing2, BAO Zhe-wei2, YANG Jing-ya1
|
|
1.Shanghai Ocean University;2.College of Food Science and Technology,Shanghai Ocean University
|
| Abstract: |
| Objective: This paper aims to study the inhibitory effect of the thermostable direct hemolysin (TDH) on mouse melanoma cell B16 in vivo and in vitro. Methods: by MTT assay, clone formation test, apoptosis test, activity test of Caspase-8 and Caspase-3, detection of mitochondrial membrane potential and tumor bearing test of C57BL / 6 mice in vivo, the half maximal inhibitory concentration IC50 of TDH on different cells were compared, and the inhibitory effect of TDH on mouse melanoma cell B16 in vivo and in vitro was evaluated. Results: the IC50 of human colon epithelial cell NCM460, human liver fibroblast LO2, human liver cancer cell SMMC-7721 and mouse melanoma cell B16 were 151μg/mL, 118μg /mL, 54μg/mL and 42μg/mL after being treated with TDH for 24h., respectively. The IC50 of normal cells is nearly higher than that of cancer cells about 2-3 times. Below the concentration of 20μg/mL, TDH could inhibit the clonal formation of B16 cells in a dose-dependent manner. 6 mg/kg TDH significantly inhibited the growth of tumor in vivo in the transplanted tumor model (P < 0.05). Flow cytometry and fluorescent reagent detection showed that: 20μg/mL TDH could induce early apoptosis of B16 cells by 19.4% and activate Caspase-8 and Caspase-3, but did not affect mitochondrial membrane potential. Conclusion: TDH has antitumor activity in vivo and in vitro. It may cause apoptosis through the apoptosis signal pathway mediated by death receptor on the cell surface, so as to play an antitumor role. |
| Key words: Marine toxins Thermostable direct hemolytic toxin Antitumor Death receptor Apoptosis |