阿托伐他汀调控DNA错配修复通路影响前列腺癌细胞放疗敏感性的机制研究

Our previous study showed that atorvastatin has radio-sensitizing effect on prostate cancer cells, but the mechanism remains unclear. This project aims to analyze the role of mismatch repair capability to radio-sensitizing effect of atorvastatin in PC3、Du145 and LNcap prostate cancer cells after radiation combined with/without atorvastatin treatment. To investigate the expression differences of Bcl-2 induced by atorvastatin and radiation combination in these three prostate cancer cell lines, and influence of radio-sensitizing effect of atorvastatin by specific interfering Bcl-2 expression, which will find the key regulatory target of radiosensitization of atorvastatin. To explore the mechanism of radioresistance of atorvastatin by mismatch repair regulation, we will detect the influence of MSH2 expression、hMutS-α mismatch bases recognizing activity and mismatch repair activity after specific interfering Bcl-2 expression; the interaction of Bcl-2 with MSH2 mismatch repair protein and subcellular localization will be investigated as well. Base on the above, we will establish prostate cancer animal model, and create the atorvastatin and radiation therapeutic schedule. The demonstration of radiosensitizing effect and its mechanism of atorvastatin in combination with radiation will enrich the theory of radiosensitization for prostate cancer and provide the experimental basis for clinical application.

我们的前期研究发现，阿托伐他汀对于前列腺癌细胞具有放疗增敏的作用，但作用机制还需进一步探讨。本研究将以PC3、Du145和LNcap细胞为模型，分别给予射线单独或联合阿托伐他汀干预，检测三种细胞错配修复能力，分析错配修复能力对于阿托伐他汀放疗增敏效应的作用。通过观察三种细胞Bcl-2的基因表达水平差异，以及特异性干扰Bcl-2表达对阿托伐他汀放疗增敏效应的影响，寻找阿托伐他汀放疗增敏的调控靶点。特异性干扰Bcl-2表达，观察MSH2表达水平、hMutS-α错配碱基识别活性和错配修复活性的影响；并通过观察Bcl-2与MSH2的蛋白质相互作用和亚细胞定位，研究错配修复通路在阿托伐他汀放疗增敏效应中的调控机制。在此基础上，构建前列腺癌荷瘤小鼠模型，并建立阿托伐他汀联合射线治疗方案，进一步明确阿托伐他汀对前列腺癌细胞的放疗增敏机制，为放疗增敏策略提供理论依据。

我们的前期研究发现，阿托伐他汀对于前列腺癌细胞具有放疗增敏的作用，但作用机制还需进一步探讨。本研究将以PC3、Du145和LNcap细胞为模型，分别给予射线单独或联合阿托伐他汀干预，检测三种细胞错配修复能力，分析错配修复能力对于阿托伐他汀放疗增敏效应的作用。通过观察三种细胞Bcl-2的基因表达水平差异，以及特异性干扰Bcl-2表达对阿托伐他汀放疗增敏效应的影响，寻找阿托伐他汀放疗增敏的调控靶点。特异性干扰Bcl-2表达，观察MSH2表达水平、hMutS-α错配碱基识别活性和错配修复活性的影响；并通过观察Bcl-2与MSH2的蛋白质相互作用和亚细胞定位，研究错配修复通路在阿托伐他汀放疗增敏效应中的调控机制。在此基础上，构建前列腺癌荷瘤小鼠模型，并建立阿托伐他汀联合射线治疗方案，进一步明确阿托伐他汀对前列腺癌细胞的放疗增敏机制，为放疗增敏策略提供理论依据。