Xu Tong, Huang Lunhua, Liu Zhiqiang, Ma Dongwen, Zhang Guowei, Ning Xiaofei, Lu Xinyang, Liu Hongsheng, Jiang Biao
Department of Gastrointestinal Surgery and 2Department of Cardiac Intensive Medicine, Affiliated Hospital of Jining Medical University, Jining, Shandong, 272000 China.
J BUON. 2019 Mar-Apr;24(2):686-692.
Totarol is a plant-derived natural product and has been reported to exhibit important pharmacological activities. However, the anticancer activity of totarol has not been evaluated yet. Therefore, the present research work was designed to evaluate the antitumor effects of totarol in SGC-7901 human gastric cancer cells and human gastric epithelial mucosa cell line GES-1 (used as normal cell line model) together with examining its effects on induction of apoptosis, cell cycle phase distribution and cell migration.
The effect of totarol on cell cytotoxicity was evaluated by MTT cell viability assay. Inverted phase contrast microscopy was used to identify the effects on cell morphology, while transmission electron microscopy indicated the apoptosis-driven morphological changes in cancer cells. The effects on cell apoptosis were also evaluated by annexin V/PI staining, while cell cycle analysis was done by flow cytometry. In vitro wound healing assay estimated the effects of totarol on cell migration.
The results indicated that totarol induced selective cytotoxic effects in SGC-7901 human gastric cancer cells concentration-dependently and exhibited lower toxicity in GES-1 normal cells. The totarol-treated cells showed significant alterations in cell morphology including rounding and cellular shrinkage. Untreated SGC-7901 cells exhibited normal cellular morphology with undamaged plasma membrane. However, treating cells with totarol led to damaged plasma membrane along with appearance of rounded protrusions (apoptotic bodies) containing damaged and broken chromatin material. Treatment with different doses of totarol led to profound suppression of wound healing. Totarol treatment also led to G2/M phase cell cycle arrest in these cells in a concentration-dependent manner.
The present study indicated that totarol diterpene has the tendency to show selective anticancer effects in SGC-7901 human gastric cancer cells along with inducing apoptosis, cell cycle arrest and inhibition of cell migration.
托塔酚是一种植物来源的天然产物，据报道具有重要的药理活性。然而，托塔酚的抗癌活性尚未得到评估。因此，本研究旨在评估托塔酚对SGC - 7901人胃癌细胞和人胃上皮黏膜细胞系GES - 1（用作正常细胞系模型）的抗肿瘤作用，并研究其对细胞凋亡诱导、细胞周期阶段分布和细胞迁移的影响。
采用MTT细胞活力测定法评估托塔酚对细胞毒性的影响。利用倒置相差显微镜观察其对细胞形态的影响，而透射电子显微镜则显示癌细胞中凋亡驱动的形态变化。通过膜联蛋白V/PI染色评估其对细胞凋亡的影响，同时采用流式细胞术进行细胞周期分析。体外伤口愈合试验评估托塔酚对细胞迁移的影响。
结果表明，托塔酚对SGC - 7901人胃癌细胞具有浓度依赖性的选择性细胞毒性作用，而对GES - 1正常细胞的毒性较低。经托塔酚处理的细胞在细胞形态上表现出显著变化，包括细胞变圆和收缩。未处理的SGC - 7901细胞呈现正常的细胞形态，细胞膜未受损。然而，用托塔酚处理细胞会导致细胞膜受损，并出现含有受损和破碎染色质物质的圆形突起（凋亡小体）。用不同剂量的托塔酚处理会导致伤口愈合受到显著抑制。托塔酚处理还会使这些细胞以浓度依赖的方式发生G2/M期细胞周期阻滞。
本研究表明，托塔酚二萜在SGC - 7901人胃癌细胞中具有显示选择性抗癌作用的趋势，同时可诱导细胞凋亡、细胞周期阻滞并抑制细胞迁移。
