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目的:探讨乙酰紫草素对人喉癌Hep-2细胞增殖和凋亡的影响及其相关的分子机制。方法:体外培养Hep-2细胞，按照处理因素不同，将实验对象分为时间处理组[以25 μmol/L(近ICn 50值)乙酰紫草素分别处理细胞0，3，6，12，24 h)]及浓度处理组(以0、10、20、30、40、50 μmol/L乙酰紫草素处理细胞24 h)，细胞增殖与毒性实验(cell counting Kit-8，CCK-8)检测不同浓度乙酰紫草素对处理不同时间人喉癌HEP-2细胞的杀伤作用；流式细胞术检测乙酰紫草素处理后Hep-2细胞凋亡情况；Western blotting检测细胞凋亡相关蛋白和Wnt/β-catenin信号通路相关蛋白的表达。n 结果:不同浓度(10～50 μmol/L)乙酰紫草素能够以时间和浓度依赖性抑制喉癌Hep-2细胞的增殖；半数抑制浓度为(26.83±2.47) μmol/L；ICn 50浓度乙酰紫草素处理Hep-2细胞3、6、12及24 h，细胞凋亡率分别为(15.25±1.74)%，(28.75±2.36)%，(36.51±3.78)%及(43.78±3.69)%，各处理组与对照组(0 h)相比，差异均具有统计学意义(n F＝43.58，n P<0.05)；乙酰紫草素能够通过上调Bax，Cyt-c，Caspase-3表达(n F＝12.67、23.47、9.52, n P值均<0.05)，下调Bcl-2，Caspase-9表达(n F＝22.29、15.62, n P<0.05)来诱导喉癌Hep-2细胞线粒体依赖性凋亡；乙酰紫草素能够降低Wnt1，β-catenin，c-myc和Cyclin D1蛋白的表达(n F＝32.67、19.57、13.88、28.14, n P值均<0.05)从而有效抑制喉癌Hep-2细胞中的Wnt/β-catenin信号通路。n 结论:乙酰紫草素能够有效抑制人喉癌Hep-2细胞增殖，并能够有效诱导Hep-2细胞发生线粒体依赖性凋亡，其具体机制可能是通过调控Wnt/β-catenin信号通路来实现的。“,”Objective:To investigate the effects of acetylshikonin on proliferation and apoptosis of human laryngeal carcinoma (Hep-2) cells and the related molecular mechanisms.Methods:Hep-2 cells were cultured in vitro, the subjects were divided into groups according to different time [25 μmol/L (near IC n 50 value) cells were treated 0, 3, 6, 12, 24 h respectively] and treatment group according to different concentrations (cells were treated by acetyl shikonin with 0, 10, 20, 30, 40, 50 μmol/L for 24 h), Cell counting kit-8 (CCK-8) was used to detect the killing effect of different concentrations of acetoshikonin on Hep-2 cells treated with different time. Flow cytometry was used to detect the apoptosis of Hep-2 cells treated with acetylshikonin. Western blotting was used to detect the expression of apoptosis-related proteins and Wnt/β-catenin signaling pathway-related proteins.n Results:Acetylshikonin can inhibit the proliferation of laryngeal cancer Hep-2 cells in a time and concentration-dependent manner, with a half inhibitory concentration of (26.83±2.47)μmol/L. The ICn 50 concentration of acetylshikonin treated Hep-2 cells for 3, 6, 12 and 24 h, the apoptosis rates were (15.25±1.74)%, (28.75±2.36)%, (36.51±3.78)% and (43.78±36.9)%, compared with the control group (0 h), the difference between each treatment group was statistically significant (n F = 43.58, n P<0.05); Acetyl shikonin can up-regulate the expression of Bax, Cyt-c, and Caspase-3 (n F=12.67, 23.47, 9.52, n all P<0.05), and down-regulate the expression of Bcl-2, Caspase-9 (n F=22.29, 15.62, both n P values <0.05) to induce mitochondrial-dependent apoptosis in laryngeal cancer Hep-2 cells; Acetyl shikonin can reduce the expression of Wnt1, β-catenin, c-myc and Cyclin D1 proteins ( n F=32.67, 19.57, 13.88, 28.14, all n P values <0.05), thereby effectively inhibiting Wnt/β-catenin signaling pathway.n Conclusion:Acetylshikonin can effectively inhibit the proliferation of human laryngeal carcinoma Hep-2 cells and induce mitochondria-dependent apoptosis in Hep-2 cells.The specific mechanism may be achieved by regulating Wnt/β-catenin signaling pathway.