白花丹素通过PI3K/AKT/mTOR通路增强PF方案杀伤舌鳞癌细胞

Plumbagin enhances the inhibition effects of PF chemotherapy regimen on tongue squamous cell carcinoma cells via PI3K/AKT/mTOR-mediated pathway

目的:研究白花丹素(plumbagin,PB)在PF方案中对舌鳞癌细胞凋亡和自噬的作用及相关机制。方法:体外培养舌癌细胞株CAL27,分为空白组、白花丹素(PB)组、顺铂+5氟尿嘧啶(PF)组、PB+PF组。MTT法检测药物对细胞的抑制作用;流式细胞仪检测细胞凋亡率;Cyto ID荧光显微镜检测细胞自噬水平;Western Blot检测细胞自噬相关蛋白Beclin1、LC3及信号通路蛋白的表达。结果:PB增强PF方案对CAL27细胞的增殖抑制作用(P<0.05);增强PF方案对CAL27细胞的凋亡率(P<0.05);增强PF方案对CAL27细胞的自噬水平(P<0.05);PB联合PF方案较单独PF方案作用,CAL27细胞中PI3K、p-AKT、p-m TOR蛋白表达均下降(P<0.05);分别应用PI3K/AKT激动剂IGF-1、m TOR激动剂MHY1485作用后,该激动剂能够逆转PB对PF方案CAL27细胞中PI3K、p-AKT、p-m TOR蛋白的下调作用(P<0.05),同时抑制了PB增强PF方案对CAL27细胞的凋亡与自噬作用。结论:白花丹素通过抑制PI3K/AKT/m TOR通路诱导凋亡和增强自噬作用从而增强PF化疗方案杀伤舌鳞癌CAL27细胞。

Objective： To investigate the effects and mechanisms of plumbagin（ PB） comebined with PF chemotherapy reginmen on apoptosis and autophagy of tongue squamous cell carcinoma cells. Methods： CAL27 cells were cultured in vitro and treated by PB,PF（ cisplatin ＋ 5-fluorouracil） and PB ＋ PF respectively. MTT assay was employed to examine the cell proliferation. Flow cytometry was used to detect the cells apoptosis. Cyto-IDautophagy detection kit was used to detect the cell autophagy level. The autophagy related proteins of Beclin1,LC3 and PI3 K/AKT/m TOR were assessed by Western Blot. Results： PB enhanced proliferation inhibition effect of PF on CAL27 cells（ P 〈 0. 05）; enhanced apoptosis rate of CAL27 cells（ P 〈 0. 05）; enhanced the autophagy level of the cells treated by PF（ P 〈 0. 05）; decreased the expression of PI3 K,p-AKT and p-m TOR（ P 〈 0. 05）. Pretreatment with IGF-I or MHY1485 inhibited the apoptosis and autophagy of PB enhanced PF chemotherapy,and reversed the down-regulation effect of PB on PI3 K,p-AKT and p-m TOR proteins expression in PF treated cells（ P 〈 0. 05）. Conclusion： Plumbagin may enhance the killing of PF chemotherapy regimen on tongue squamous cell carcinoma cells by inducing apoptosis and autophagy via inhibiting PI3 K/AKT/m TOR mediated pathway.