红景天苷对人舌癌细胞的凋亡作用及机制研究

目的：探讨红景天苷对人舌癌细胞增殖和凋亡的影响，并解释其分子机制，为舌癌的临床治疗提供新的思路。方法：CCK-8法和克隆形成实验检测红景天苷药物对人舌癌细胞增殖能力的影响；Annexin V/PI双染法和TUNEL法检测细胞凋亡情况；PI染色法检测红景天苷对细胞周期的影响；细胞迁移法检测红景天苷对舌癌细胞迁移的影响；免疫印迹法和实时荧光定量PCR法检测细胞内凋亡相关蛋白Bax、Bcl-2和Caspase 3的表达水平；免疫印迹法检测细胞内ERK和PI3K/AKT信号通路关键蛋白的表达水平的变化。结果：红景天苷能显著抑制人舌癌细胞的增殖且呈剂量依赖性。Annexin V/PI双染法和TUNEL法显示，随着红景天苷浓度的增加，Tca-8113细胞凋亡明显增多。PI染色法显示，红景天苷处理细胞后，G2/M期细胞明显减少。细胞迁移实验表明，红景天苷作用于舌癌细胞后，随着给药浓度的逐渐升高，细胞的迁移能力明显降低。免疫印迹结果表明，红景天苷可以使Bax和Cleaved caspase 3蛋白表达水平升高，Bcl2的蛋白表达水平降低。同时，红景天苷可以抑制ERK和PI3K/AKT信号通路。结论：红景天苷可以明显抑制人舌癌Tca-8113细胞的增殖，诱导其凋亡，其作用机制可能与抑制ERK和PI3K/AKT信号通路有关。

Pro-apoptotic mechanism of salidroside for human tongue cancer cell

OBJECTIVE To explore the effect of salidroside on the proliferation and apoptosis of human tongue cancer cells, elucidate its molecular mechanism and provide new ideas for clinical therapeutics of tongue cancer.METHODS CCK-8 and colony formation assays were utilized for detecting the effect of salidroside on cell proliferation; Annexin V/PI staining and TUNEL assay for detecting cellular apoptosis; PI staining assay for detecting the effect of salidroside on cell cycle; Cell migration assay for detecting the effect of salidroside on the migration of tongue cancer cells. The protein expression levels of Bax, Bcl-2 and Caspase 3 were detected by Western blot and real-time polymerase chain reaction (PCR). And Western blot was employed for detecting the key protein expression levels of ERK and PI3K/AKT signaling pathways.RESULTS Salidroside significantly suppressed the proliferation of human tongue cancer cells in a dose-dependent manner. Annexin V/PI double staining and TUNEL assays indicated that apoptosis of Tca-8113 cell spiked markedly with a rising drug concentration. PI staining showed that the number of cells in G2/M phase declined significantly after salidroside treatment. Cell migration experiments revealed that the migratory capability of tongue cancer cells decreased markedly with a rising concentration of salidroside. Western blot denoted that salidroside could boost the protein expression levels of Bax and Cleaved caspase 3 and lower the protein expression level of Bcl-2. At the same time, salidroside could suppress the signaling pathways of ERK and PI3K/AKT.CONCLUSION Salidroside can significantly suppress the proliferation of Tca-8113 cell and induce cellular apoptosis. The mechanism of action may be correlated with the suppressions of ERK and PI3K/AKT signaling pathways.