Hong Bin Wu, Da Lu Hong, Xue Yin, Jing BaoWen
Department of Oncology, the Central Hospital of Wuhan, Tongji Medical College， Huazhong University of Science and Technology, Wuhan, Hubei 430014, China.
J BUON. 2018 Sep-Oct;23(5):1384-1389.
To examine the molecular mechanism of action behind the anticancer effects of pterostilbene in HeLa human cervical cancer cells.
MTS assay was used to study the pterostilbene cytotoxic effects, while inverted phase contrast and fluorescence microscopy was used to study the effects of the drug on the cell apoptosis and changes in cell morphology. Flow cytometry was used to study changes in mitochondrial membrane potential (MMP), while immunoblotting assay was used to demonstrate its effects on m-TOR/PI3K/Akt protein signalling pathway.
Pterostilbene induced potent, dose-dependent and time-dependent cytotoxic effects in HeLa cancer cells exhibiting IC50 of 101.2 µM and 65.9 µM at 24 and 48 hrs time intervals, respectively. As compared to the untreated control cells which revealed normal cell morphology, pterostilbene-treated cells exhibited significant cellular shrinkage which increased with increasing doses of the drug. Untreated control cells showed complete green fluorescence corresponding to absence of apoptosis. However, pterostilbene-treated cells with 25, 100 and 200 µM showed increasing emission of red/orange fluorescence corresponding to apoptotic induction. Pterostilbene-treated cells showed evident signs of DNA ladder formation and the effect increased with increasing concentrations of the drug. The percentage of cells with depolarized mitochondria (loss of MMP) increased from 2.3% in the control group to 24.5, 43.2 and 65.8% in cells treated with 0, 25, 100 and 200 µM concentration of pterostilbene, respectively.
Pterostilbene exerts potent anticancer effects in HeLa human cervical cancer cells via disruption of MMP, apoptosis induction and targeting m-TOR/PI3K/Akt signalling pathway.
研究白藜芦醇对人宫颈癌HeLa细胞抗癌作用背后的分子作用机制。
采用MTS法研究白藜芦醇的细胞毒性作用，同时利用倒置相差显微镜和荧光显微镜研究该药物对细胞凋亡及细胞形态变化的影响。采用流式细胞术研究线粒体膜电位（MMP）的变化，同时利用免疫印迹法证明其对m-TOR/PI3K/Akt蛋白信号通路的影响。
白藜芦醇对HeLa癌细胞具有显著的、剂量依赖性和时间依赖性的细胞毒性作用，在24小时和48小时间隔时的半数抑制浓度（IC50）分别为101.2 μM和65.9 μM。与显示正常细胞形态的未处理对照细胞相比，经白藜芦醇处理的细胞表现出明显的细胞皱缩，且随着药物剂量的增加而加剧。未处理的对照细胞显示出完整的绿色荧光，表明不存在凋亡。然而，用25、100和200 μM白藜芦醇处理的细胞显示出红色/橙色荧光发射增加，表明诱导了凋亡。经白藜芦醇处理的细胞显示出明显的DNA梯形条带形成迹象，且该效应随着药物浓度的增加而增强。线粒体去极化（MMP丧失）的细胞百分比从对照组的2.3%分别增加到用0、25、100和200 μM浓度白藜芦醇处理的细胞中的24.5%、43.2%和65.8%。
白藜芦醇通过破坏MMP、诱导凋亡和靶向m-TOR/PI3K/Akt信号通路，对人宫颈癌HeLa细胞发挥强大的抗癌作用。
