Wang Ziqian, Tang Tuo, Wang Shengnan, Cai Tianyu, Tao Hong, Zhang Qing, Qi Shimei, Qi Zhilin
Department of Biochemistry and Molecular Biology, Wannan Medical College, Wuhu, Anhui 241002, People's Republic of China.
Anhui Province Key Laboratory of Active Biological Macro-Molecules, Wannan Medical College, Wuhu, Anhui 241002, People's Republic of China.
Drug Des Devel Ther. 2020 Jan 14;14:145-155. doi: 10.2147/DDDT.S219247. eCollection 2020.
Aloin has been reported to have many pharmacological effects including anti-inflammatory, anti-oxidant and anti-tumour activities. However, the precise molecular mechanisms underlying the anti-tumour properties of aloin are yet to be elucidated.
HGC-27 and BGC-823 gastric cancer cells were treated with aloin. EdU and colony formation assays were used to detect the proliferation ability of cells. The migration of cells was detected using wound healing and transwell assays. Western blotting was used to detect the levels of cyclinD1, cyclin E1, MMPs, N-cadherin, E-cadherin and NOX2. The phosphorylation of Akt, mTOR, P70S6K, S6, Src, stat3 and IκBα were also detected by Western blotting. Flow cytometry was used to detect the cell cycle distribution.The location of p65 in cells was determined by using a confocal microscopy assay. The total amounts of ROS present in cells were measured using an ROS assay kit.
Here, we found that aloin inhibited the proliferation and migration of HGC-27 and BGC-823 gastric cancer cells using a combination of EdU, colony formation, wound healing and transwell assays. Further investigations revealed that aloin decreased the protein expression levels of cyclin D1, N-cadherin, and the matrix metalloproteinases (MMP)-2 and MMP-9; increased E-cadherin expression in a dose-dependent manner; inhibited reactive oxygen species (ROS) generation; and mediated the activation of Akt-mTOR, signal transducer and activator of transcription-3 (Stat3), and NF-κB signalling pathways. Our results also indicated that aloin is able to attenuate the expression levels of the two regulatory proteins of nicotinamide adenine dinucleotide phosphate oxidase 2 (NOX2), p47 and p22, but had no effect on the level of gp91. N-acetylcysteine treatment of gastric cancer cells inhibited ROS production and Akt-mTOR, Stat3, and IκBα phosphorylation. Taken together, our data suggest that aloin inhibits the proliferation and migration of gastric cancer cells by downregulating NOX2-ROS-mediated activation of the Akt-mTOR, Stat3, and NF-κB signalling pathways.
Our findings suggest a potential role for aloin in the prevention of gastric cancer cell proliferation and migration and provide novel insights into the anti-cancer properties of aloin.
据报道，芦荟素具有多种药理作用，包括抗炎、抗氧化和抗肿瘤活性。然而，芦荟素抗肿瘤特性的精确分子机制尚待阐明。
用芦荟素处理人胃癌细胞系HGC-27和BGC-823。采用EdU和集落形成试验检测细胞增殖能力。采用划痕愈合试验和Transwell试验检测细胞迁移情况。采用蛋白质免疫印迹法检测细胞周期蛋白D1、细胞周期蛋白E1、基质金属蛋白酶（MMP）、N-钙黏蛋白、E-钙黏蛋白和NOX2的水平。同时采用蛋白质免疫印迹法检测Akt、mTOR、P70S6K、S6、Src、信号转导子和转录激活子3（Stat3）以及IκBα的磷酸化水平。采用流式细胞术检测细胞周期分布。采用共聚焦显微镜检测p65在细胞内的定位。使用活性氧（ROS）检测试剂盒测定细胞内ROS的总量。
在此，我们发现，通过EdU、集落形成、划痕愈合和Transwell试验联用，芦荟素可抑制HGC-27和BGC-823胃癌细胞的增殖和迁移。进一步研究表明，芦荟素可降低细胞周期蛋白D1、N-钙黏蛋白以及基质金属蛋白酶-2（MMP-2）和基质金属蛋白酶-9（MMP-9）的蛋白表达水平；以剂量依赖性方式增加E-钙黏蛋白的表达；抑制活性氧生成；并介导Akt-mTOR、信号转导子和转录激活子3（Stat3）以及核因子κB（NF-κB）信号通路的激活。我们的结果还表明，芦荟素能够降低烟酰胺腺嘌呤二核苷酸磷酸氧化酶2（NOX2）的两种调节蛋白p47和p22的表达水平，但对gp91的水平无影响。用N-乙酰半胱氨酸处理胃癌细胞可抑制ROS的产生以及Akt-mTOR、Stat3和IκBα的磷酸化。综上所述，我们的数据表明，芦荟素通过下调NOX2-ROS介导的Akt-mTOR、Stat3和NF-κB信号通路的激活来抑制胃癌细胞的增殖和迁移。
我们的研究结果表明芦荟素在预防胃癌细胞增殖和迁移方面具有潜在作用，并为芦荟素的抗癌特性提供了新的见解。
