| 摘要: |
| 为探讨耐热直接溶血毒素(Thermostabile direct hemolysin,TDH)在体内和体外对小鼠黑色素瘤细胞B16的抑制作用,本研究通过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(Mus musculus)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。结果发现:人结肠上皮细胞NCM460、人正常肝细胞LO2、人肝癌细胞SMMC-7721和小鼠黑色素瘤细胞B16在TDH处理24 h之后,细胞的半抑制质量浓度IC50分别为151、118、54和48 μg/mL,正常细胞的IC50高出癌细胞近2~3倍。当质量浓度低于20 μg/mL时,TDH以剂量依赖性的方式抑制B16细胞的克隆形成,6 mg/kg TDH在移植瘤模型中显著抑制体内肿瘤的生长(P<0.001)。流式细胞术和荧光试剂盒检测表明:20 μg/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。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：10.11759/hykx20220328006 |
| 分类号:R979.1 |
| 基金项目:上海市教委高原学科中青年人才培育项目(A1-3203-20-100617) |
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| Study on the inhibitory effect of thermostable direct hemolytic toxin from Vibrio parahaemolyticus on melanoma |
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YANG Lan-zhu, LI Shi-yi, ZHENG Wen-jing, LI Jing, BAO Zhe-wei, YANG Jing-ya
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College of Food Science and Technology, Shanghai Ocean University, Shanghai 201306, China
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| Abstract: |
| This paper aims to investigate the inhibitory effect of thermostable direct hemolysin (TDH) on B16 mouse melanoma cells in vivo and in vitro. The half maximal inhibitory concentration (IC50) of TDH on different cells were compared, and the inhibitory effect of TDH on B16 mouse melanoma cells in vivo and in vitro was evaluated using 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. After 24 h of TDH treatment, the IC50 of human colon epithelial cell NCM460, human liver fibroblast LO2, human liver cancer cell SMMC-7721, and mouse melanoma cell B16 were 151 μg/mL, 118 μg/mL, 54 μg/mL, and 42 μg/mL, respectively. The IC50 of normal cells was 2–3 times higher than that of cancer cells. TDH could inhibit the clonal formation of B16 cells in a dose-dependent manner below a concentration of 20 μg/mL. TDH at 6 mg/kg significantly inhibited tumor growth in vivo in the transplanted tumor model (P<0.05). Flow cytometry and fluorescent reagent detection revealed that 20 μg/mL TDH could induce early apoptosis in B16 cells by 19.4% and activate caspase-8 and caspase-3, but it had no effect on mitochondrial membrane potential. TDH has antitumor activity both in vivo and in vitro. It may induce apoptosis via the apoptosis signal pathway mediated by the death receptor on the cell surface, thereby acting as an antitumor agent. |
| Key words: Marine toxins thermostable direct hemolytic toxin antitumor death receptor apoptosis |
