LY294002联合顺铂通过细胞自噬效应抑制人喉癌HEP-2细胞的生长

LY294002 combined with cisplatin inhibits the growth of human laryngeal carcinoma HEP-2 cells by autophagy

目的:探讨PI3K/AKT通路抑制剂LY294002联合顺铂抑制人喉癌HEP-2细胞生长的机制。方法:采用MTT及细胞活力检测单独LY294002处理对HEP-2细胞的毒性作用,并且观察LY294002联合顺铂对HEP-2细胞产生的影响;采用免疫荧光检测LY294002联合顺铂对HEP-2细胞内自噬特异性标志蛋白LC3B表达的诱导作用;采用实时荧光定量PCR(RT-qPCR)检测LY294002联合顺铂处理HEP-2细胞后相关自噬因子的表达。结果:单独LY294002对喉癌HEP-2细胞无明显毒性作用,而1μg/ml顺铂联合2μg/ml LY294002可显著增强顺铂杀伤HEP-2细胞的效能,并且明显刺激LC3B的表达。RT-qPCR结果显示,1μg/ml顺铂联合2μg/ml LY294002作用显著升高细胞中自噬标志物ATG3、ATG4A、ATG4B、ATG4C、ATG5和ATG7 mRNA的表达。结论:顺铂联合LY294002可能通过激活细胞自噬抑制喉癌HEP-2细胞生长的作用。

Objective:To investigate the mechanism of the inhibitory effect of LY294002 combined with cisplatin on the growth of human laryngeal carcinoma HEP-2 cells.Methods:MTT and cell viability were used to detect the cytotoxic effect of LY294002 alone and LY294002 combined with cisplatin on HEP-2 cells.The effect of LY294002 combined with cisplatin on autophagic marker LC3B in HEP-2 cells was detected by immunofluorescence.And the expressions of autophagic factors in autophagy pathway were detected by real-time quantitative PCR(RT-qPCR).Results:LY294002 alone had no significant cytotoxic effect on HEP-2 cells,while 2μg/ml LY294002 combined with 1μg/ml cisplatin significantly enhanced the killing effect of cisplatin on the cells,and greatly increased the expression of LC3B.The results of RT-qPCR showed that LY294002 combined with cisplatin significantly increased the mRNA expression of ATG3,ATG4A,ATG4B,ATG4C,ATG5 and ATG7.Conclusion:The combination of LY294002 and cisplatin may inhibit the growth of HEP-2 cells by activating autophagy.