橙皮苷对人舌癌Tca8113细胞增殖和凋亡的影响及机制研究

Effect and mechanism of hesperidin on proliferation and apoptosis of human tongue cancer Tca8113 cells

目的:探讨橙皮苷(HP)对人舌癌Tca8113细胞增殖和凋亡的影响及机制。方法:将人舌癌Tca8113细胞分为对照组、低剂量HP组和高剂量HP组。对照组细胞常规培养。低剂量HP组细胞在对照组基础上给予30μmol/L HP,高剂量HP组细胞则给予60μmol/L HP。24 h后,采用CCK-8实验检测细胞增殖活力;采用Hoechst 33342染色和Annexin V-FITC/PI染色检测细胞凋亡情况;采用Western blot检测凋亡相关蛋白磷酸化磷脂酰肌醇3-激酶(p-PI3K)、磷酸化蛋白激酶B(p-AKT)、B细胞淋巴瘤/白血病-2(Bcl-2)和Bcl-2相关X蛋白(Bax)的表达情况。结果:经30、60μmol/L HP处理24 h后,舌癌Tca8113细胞的增殖活力受到不同程度的抑制(均P<0.01)。对照组细胞形态规则,染色质均匀,胞核呈现出弱蓝色荧光;HP实验组细胞染色质浓缩,细胞核固缩,部分胞核碎裂呈现出亮蓝色荧光,凋亡数量明显增加。HP实验组细胞凋亡率高于对照组(均P<0.01)。低剂量HP组和高剂量HP组细胞p-PI3K、p-AKT和Bcl-2蛋白相对表达量明显低于对照组,而Bax蛋白相对表达量显著高于对照组(均P<0.01)。结论:HP可通过调控PI3K/AKT信号通路抑制人舌癌Tca8113细胞增殖活力,促进凋亡。

Objective:To investigate the effect and mechanisms of Hesperidin(HP)on proliferation and apoptosis of human tongue cancer Tca8113 cells.Methods:Human tongue cancer Tca8113 cells were divided into control group,low-dose HP group and high-dose HP group.Cells in the control group were routinely cultured.Cells in the low-dose HP group received 30μmol/L HP on the basis of the control group,while cells in the high-dose HP group received 60μmol/L HP.After 24 hours,cell proliferation activity was measured by CCK-8 assay;apoptosis was detected by Hoechst33342 staining and Annexin V-FITC/PI staining;and expression of apoptosis-related proteins p-PI3 K,p-AKT,Bcl-2 and Bax were measured by Western blot.Results:After 30 and 60μmol/L HP treatment for 24 hours,the proliferation of tongue cancer Tca8113 cells was inhibited to different degrees(all P<0.01).In the control group,the cells had regular morphology,uniform chromatin,and weak blue fluorescence in the nucleus;in the HP experimental group,the chromatin was condensed,the nucleus was pyknotic,and some nuclei were fragmented,showing bright blue fluorescence,and the number of apoptotic cells increased significantly.The apoptosis rate of HP experimental group was higher than that of control group(all P<0.01).The relative expressions of p-PI3K,p-AKT and Bcl-2 proteins in the low-dose HP group and the high-dose HP group were significantly lower than those in the control group,while the relative expression of Bax protein was significantly higher than that in the control group(all P<0.01).Conclusion:Hesperidin can inhibit the proliferation of human tongue cancer Tca8113 cells and promote apoptosis by regulating PI3K/AKT signaling pathway.