植物化学物质的抗癌机制是备受关注的研究热点。我们近期研究发现，薄荷醇能够通过差异性操控癌细胞和正常细胞内质网钙离子释放，显著提高木犀草素的抗癌特异性。但是，导致这一现象的分子机制，目前仍不清楚。因此，本项目以3种癌细胞和3种正常细胞为模型，采用细胞钙离子示踪、膜通道封堵、细胞器分离以及siRNA技术，研究IP3R、RyR、TRPV1以及TRPM8钙离子通道在癌/正常细胞内质网膜上的表达差异，揭示薄荷醇差异性操控癌/正常细胞内质网钙离子释放的分子机制；通过酶活性分析实验，考察钙离子对NADPH-醌氧化还原酶（PIG3）活性的影响，明确PIG3对薄荷醇提高木犀草素抗癌特异性的介导作用；采用半定量RT-PCR、流式细胞及免疫印迹技术，确定薄荷醇协同木犀草素诱导癌细胞凋亡时内质网应激和线粒体损伤信号的传递过程。本研究对于深入认识植物化学物质之间的协同抗癌机制，具有重要的科学理论意义。

Anticancer mechanism of phytochemicals has become a research focus. In recent study, we found that the anticancer specificity of luteolin was enhanced markedly by Ca2+ release from endoplasmic reticulum which was manipulated differently by menthol in cancer cells and normal cells. However, the molecular mechanism leading to this phenomenon is unclear by now. Therefore, in this project, the expressions of IP3R, RyR, TRPV1, and TRPM8 on endoplasmic reticulum membrane will be studied in three cancer cell lines and three normal cell lines by using Ca2+ indicator, membrane channel inhibitor, subcellular separation and siRNA, to reveal the molecular mechanism on difference of Ca2+ release manipulated by menthol from endoplasmic reticulum in cancer cells and normal cells. The effect of Ca2+ on a NADPH-quinone oxidoreductase (PIG3) will be investigated by enzyme activity analysis, to make clear the mediating role of PIG3 between menthol and the anticancer specificity of luteolin. The apoptotic signal pathways of mitochondria damage and endoplasmic reticulum stress induced by menthol and luteolin in cancer cells will be determined by semi-quantitative RT-PCR, flow cytometry and western blot. This study has a scientific theoretical significance in understanding the synergistic anticancer mechanism between phytochemicals.

植物化学物质的抗癌机制是备受关注的研究热点。我们前期研究发现，薄荷醇能够显著提高木犀草素的抗癌特异性。但是，导致这一现象的分子机制，目前仍不清楚。本项目以3种癌细胞和3种正常细胞为模型，采用免疫荧光、细胞钙离子示踪、膜通道封堵、细胞器分离以及Western-blot等技术，研究IP3R、RyR、TRPV1以及TRPM8钙离子通道在癌细胞与正常细胞内质网上的表达差异，揭示薄荷醇差异性操控癌细胞与正常细胞内质网钙离子释放的分子机制。研究取得了以下结果：（1）薄荷醇对木犀草素抗癌特异性的促进作用与其差异性操控癌细胞与正常细胞内质网钙离子释放有关；（2）TRPM8在癌细胞中表达明显高于正常细胞，而IP3R、RyR、TRPV1在癌细胞与正常细胞中表达无明显差异性，因此TRPM8是导致薄荷醇区别操控癌细胞与正常细胞内质网钙离子释放的关键因素；（3）低温有利于薄荷醇对细胞内质网钙离子释放活性的发挥；（4）IP3R除定位于细胞内质网外，也存在于细胞核，其功能目前尚不清楚，具有后续研究价值。上述研究结果对于深入认识植物化学物质之间的协同抗癌机制，具有重要的科学理论意义。