| 引用本文： |
-
吴强,桂亚平,袁涛,等.SENL通过mTOR通路抑制前列腺癌的研究[J].同济大学学报（医学版）,2016,37(4):40-45. [点击复制]
- WU Qiang,GUI Ya-ping,YUAN Tao,et al.SENL inhibits prostate carcinoma cells via mTOR pathway[J].同济大学学报（医学版）,2016,37(4):40-45. [点击复制]
|
|
| 摘要: |
| 目的探讨印楝(Azadirachta Indica)抑制前列腺癌进展的分子机制。方法 利用超临界流体萃取技术,制备SENL。不同浓度SENL处理人前列腺癌LNCaP-luc2、PC3细胞24h,细胞计数法和MTS法检测SENL对LNCaP-luc2、PC3细胞生长曲线的影响并计算增殖抑制率；观察细胞形态学改变；AnnexinV法检测早期凋亡率；免疫细胞化学法检测癌细胞增殖、凋亡及侵袭相关蛋白mTOR、s6K70、integrin、FAK、AR等的表达差异,免疫荧光法检测侵袭相关的蛋白表达差异。结果 SENL能抑制前列腺癌细胞的增殖,且成剂量依赖性。经IC50处理的前列腺癌细胞,免疫印迹法检测到mTOR通路的核心成分mTOR、s6K70蛋白的表达显著受到抑制,同时integrin、FAK水平明显下调。结论 体外实验中证实SENL通过抑制mTOR及相关通路活性,显著抑制前列腺癌细胞的增殖,诱导细胞凋亡。 |
| 关键词: 印楝超临界萃取物 mTOR通路 前列腺肿瘤 |
| DOI：10.16118/j.1008-0392.2016.04.008 |
| 投稿时间：2015-11-16 |
| 基金项目: |
|
| SENL inhibits prostate carcinoma cells via mTOR pathway |
| WU Qiang,GUI Ya-ping,YUAN Tao,BIAN Cui-dong,WU Deng-long |
| (Dept. of Urology, Tongji Hospital, Tongji University, Shanghai 200065, China) |
| Abstract: |
| Objective To investigate the effects of supercritical extract of Azadirachta Indica(neem) leaves (SENL) on proliferation of prostate cancer cells and its mechanism. Methods Supercritical extract of fresh neem leaves was prepared using the Speed SFE-2 System. Androgen dependent prostate adenocarcinoma LNCaP-luc2-cells and androgen independent PC3 cells were treated with different concentrations of SENL(25,0,12.5,0,6.25,5,0mg/ml) for 24 hours. Proliferation of LNCap-luc2 and PC3 cells were determined by MTS-formazan reduction method. LNCaP-luc2 and PC3 cells were treated with IC50 concentrations of SENL (12 or 15mg/ml, respectively) and stained with annexin-V FITC and propidium iodide (PI), then analyzed by flow-cytometry. The expressions of mTOR, s6K70, integrin, FAK, androgen receptor were dected by Western blotting. LNCaP-luc2 and PC3 cells were stained with immunofluorescence and observed under confocal microscope. Results SENL exhibited growth of LNCaP-luc2 and PC3 cell in a dose-dependent manner. SENL induces apoptosis of LNCaP-luc2 and PC3 cells. Western blotting analysis showed that the levels of mTOR, S6K70,androgen receptor, integrin β1 and phosphorylation of mTOR, S6K70, FAK were reduced with SENL treatment. Immunofluorescence for integrin β1 and FAK further demonstrated the suppression of formation of the focal adhesions in LNCaP-luc2 and PC3 cells after SENL treatment. Conclusion Supercritical extract of Azadirachta Indica (Neem) leaves can inhibit proliferation and induce apoptosis of prostate cancer cells in vitro, in which mTOR pathway may be involved. |
| Key words: SENL mTOR prostate cancer |
