水飞蓟宾干预调控前列腺癌干细胞样细胞的作用及分子机制研究

Silibinin has pleiotropic anticancer treatment ability, and the prostate cancer has an inhibitory effect. The bioinformatics method combined with PubChem database and Subpathway-GM platform was used to analyze the new targets of silybin, and it was found that it was highly correlated with prostate cancer stem cell marker ALDH1A1. ALDH1A1 can catalyze the formation of retinoic acid (RA) by retinol, and the binding of RA and its receptor RAR activates the transcription factor Ets1, which promotes cell growth and invasion. On the basis of literature and previous research, the relationship between ALDH1A1 and RAR and Ets1 was observed in vitro and in vitro experiments. The ALDH1A1+ and ALDH1A1 prostate cancer cells were sorted by magnetic bead sorting, flow cytometry and other techniques. Comparison of the tumorigenic ability of ALDH1A1+ and ALDH1A1- prostate cancer cells in nude mice and the effects of silybin on them. To elucidate the inhibitory effect of silybin on the growth and invasion of ALDH1A1+ prostate cancer stem cell-like cells and the related molecular mechanisms. This topic reveals the molecular mechanism of silybin inhibiting the growth and invasion of prostate cancer cells from a new vision, providing new ideas and new targets for the prevention and treatment of prostate cancer.

水飞蓟宾具有多效性抗癌能力，并且对前列腺癌具有抑制作用。本项目组采用生物信息学方法，结合PubChem数据库和Subpathway-GM平台分析水飞蓟宾新的靶点，发现其与前列腺癌干细胞标志物ALDH1A1高度相关。ALDH1A1能催化视黄醛生成维甲酸（RA），而RA与其受体RAR结合能激活转录因子Ets1，促进细胞生长和侵袭。在文献和前期研究基础上，本项目通过体内外实验观察ALDH1A1与RAR、Ets1相关性；磁珠分选、流式等技术分选出ALDH1A1+和ALDH1A1-前列腺癌细胞，比较ALDH1A1+和ALDH1A1-前列腺癌细胞裸鼠致瘤能力及水飞蓟宾对其的影响；阐释水飞蓟宾调控ALDH1A1+前列腺癌干细胞样细胞生长侵袭的抑制作用及相关分子机制。本课题从新的视觉揭示水飞蓟宾抑制前列腺癌细胞生长侵袭的分子机制，为前列腺癌的防治提供新思路和新靶点。

目的意义：.基于生物信息学方法进行水飞蓟宾药物靶点筛选，旨在探究水飞蓟宾是否通过抑制ALDH1A1的表达进而抑制前列腺癌干细胞样细胞的生长，并揭示其潜在的分子机制。.主要研究内容：.1.阐明在ALDH1A1+前列腺癌干细胞样细胞中，水飞蓟宾抑制其增殖、侵袭的机制与抑制前列腺癌干细胞标志物ALDH1A1+有关。揭示ALDH1A1通过激活RAR蛋白，进一步激活Ets1，MMP9，从而促进前列腺癌的侵袭和转移。.首先利用生物信息学方法筛选出与水飞蓟宾危险代谢途径亚途径高度相关的前列腺癌干细胞标志物ALDH1A1。通过干扰ALDH1A1基因，发现ALDH1A1表达与Ets1呈正相关。重要的是，通过MTT法、transwell法、平板克隆法和球形形成法，发现ALDH1A1+细胞具有增殖、迁移和侵袭能力。此外，体外实验表明，水飞蓟宾通过抑制前列腺干细胞的增殖、迁移和侵袭能力，对前列腺干细胞有抑制作用。.2. 在NOD/SCID裸鼠体内进一步揭示水飞蓟宾通过阻断ALDH1A1表达实现对前列腺癌干细胞样细胞的抑制作用。.为了进一步证实水飞蓟宾的抗肿瘤作用，我们构建了移植性肉瘤小鼠模型。结果显示，对照组肿瘤体积在第10天后大于水飞蓟宾组，并且结果一直维持到小鼠死亡。另外，对照组的肿瘤重量比水飞蓟宾组重。通过构建移植性肉瘤小鼠模型证实了水飞蓟宾能够抑制ALDH1A1+前列腺癌干细胞的成瘤能力。并且根据Western blot发现水飞蓟宾能降低肿瘤组织内ALDH1A1、Ets1、RARα和MMP9的表达水平。.结论：.1.在ALDH1A1+前列腺癌干细胞样细胞中，水飞蓟宾抑制其增殖、侵袭的机制与抑制前列腺癌干细胞标志物ALDH1A1有关。.2. ALDH1A1通过激活RARα蛋白，进一步激活Ets1，MMP9，从而促进前列腺癌的侵袭和转移。 .3. 在NOD/SCID裸鼠体内，水飞蓟宾通过阻断ALDH1A1表达实现对前列腺癌干细胞样细胞的抑制作用。.成果共发表SCI收录论文8篇，其中第一作者（含共同）4篇，通讯作者1篇，基金标注5篇。获省政府三等奖一项，省中医药管理局一等奖1项。出版著作2部。