Yang Guang, Yin Xuelian, Ma Dongjie, Su Zhejun
Department of Stomatology,Affiliated Hospital of Chengde Medical University, Chengde 067000, China.
J BUON. 2020 Jan-Feb;25(1):344-349.
Phloretin is one of the important polyphenolics abundantly present across the plant kingdom. Studies have reported the anticancer effects of Phloretin against different human cancer cells. Nonetheless, the anticancer effects of Phloretin have not been explored against the human oral cancer cells. Therefore this study was designed to investigate the anticancer effects of Phloretin against the human oral cancer cells.
CCK-8 assay was used for the determination of cell viability. Annexin V/propidium iodide (PI) staining and flow cytometry were used for necrosis detection and cell cycle analysis, respectively. Wound healing assay was used for cell migration analysis. Western blot analysis was used for protein expression analysis.
The results showed that Phloretin suppressed the proliferation rate of the human SCC-1 oral cancer cells and showed an IC50 of 12.5 µM. Nonetheless, Phloretin had negligible effects on the proliferation rate of the EBTr normal oral cells. DAPI staining showed that Phloretin did not induce apoptosis and western blot showed that it had no apparent effects on the Bax and Bcl-2 expression. Nonetheless annexin V/PI staining showed that Phloretin caused cell death in SCC-1 oral cancer cells. Flow cytometric analysis showed that Phloretin caused increase in the reactive oxygen species (ROS) levels of the SCC-1 cells in a time and dose-dependent manner. Cell cycle analysis showed that Phloretin caused increase in the percentage of the SCC-1 cells in the G0/G1 phase of the cell cycle leading to G0/G1 cell cycle arrest. The G0/G1 arrest of SCC-1 cells was also associated with depletion of cyclin D1, CDK4 and CDK6 expression. Wound healing assay was also performed which showed that Phloretin suppressed the migration of the SCC-1 oral cancer cells, indicative of the anti-metastatic potential of Phloretin.
Phloretin exhibits significant growth inhibitory effects on the human oral cancer cells and may prove beneficial in oral cancer treatment.
根皮素是植物界中大量存在的重要多酚类物质之一。研究报道了根皮素对不同人类癌细胞的抗癌作用。然而，根皮素对人类口腔癌细胞的抗癌作用尚未得到研究。因此，本研究旨在探讨根皮素对人类口腔癌细胞的抗癌作用。
采用CCK-8法测定细胞活力。分别采用膜联蛋白V/碘化丙啶（PI）染色和流式细胞术进行坏死检测和细胞周期分析。采用伤口愈合试验进行细胞迁移分析。采用蛋白质印迹分析进行蛋白质表达分析。
结果表明，根皮素抑制了人类SCC-1口腔癌细胞的增殖率，IC50为12.5 μM。然而，根皮素对EBTr正常口腔细胞的增殖率影响可忽略不计。DAPI染色显示根皮素未诱导细胞凋亡，蛋白质印迹显示其对Bax和Bcl-2表达无明显影响。然而，膜联蛋白V/PI染色显示根皮素可导致SCC-1口腔癌细胞死亡。流式细胞术分析显示，根皮素以时间和剂量依赖性方式导致SCC-1细胞中活性氧（ROS）水平升高。细胞周期分析显示，根皮素导致SCC-1细胞在细胞周期G0/G1期的百分比增加，导致G0/G1期细胞周期停滞。SCC-1细胞的G0/G1期停滞也与细胞周期蛋白D1、细胞周期蛋白依赖性激酶4（CDK4）和细胞周期蛋白依赖性激酶6（CDK6）表达的减少有关。还进行了伤口愈合试验，结果显示根皮素抑制了SCC-1口腔癌细胞的迁移，表明根皮素具有抗转移潜力。
根皮素对人类口腔癌细胞具有显著的生长抑制作用，可能对口腔癌治疗有益。
