姜黄挥发油对皮肤鳞癌A431细胞增殖及凋亡的影响

为研究姜黄挥发油(turmeric volatile oil,TVO)对皮肤鳞癌(CSCC)A431(以下简称A431)细胞增殖及凋亡的影响并探讨其机制,该实验采用不同质量浓度(5~80 mg·L-1)TVO体外作用于A431细胞,利用细胞增殖/毒性检测试剂盒(celcounting kit-8,CCK-8)法检测细胞增殖活性;倒置显微镜观察细胞形态变化;吖啶橙(AO)/溴化乙锭(EB)双染色荧光显微镜观察A431细胞凋亡情况;流式细胞仪检测细胞凋亡;Western blot法检测半胱氨酸天冬氨酸蛋白酶-3(caspase-3)、半胱氨酸天冬氨酸蛋白酶-9(caspase-9)表达量。结果显示,随TVO质量浓度的增加对A431细胞生长具有显著的抑制增殖作用,呈剂量依赖关系,各组间差异有统计学意义(P0.05)。与对照组比较,不同质量浓度TVO组均出现细胞皱缩及细胞破碎现象,细胞凋亡率增加,并呈剂量依赖性,caspase-3,caspase-9的相对表达量上调,各组间差异有统计学意义(P0.05),提示了TVO能抑制皮肤鳞癌A431细胞增殖,并诱导其凋亡,其机制可能与上调caspase-3,caspase-9的表达有关。

Effect of turmeric volatile oil on proliferation and apoptosis of human skin SCC A431 cells

To investigate the effect of turmeric volatile oil (TVO) on the apoptosis and proliferation of human skin SCC A431 cells, A431 cells were incubated with different concentrations(5-80 mg·L^-1) of TVO in vitro.The proliferation and cell cycle were assessed by CCK8 assay. The change of morphology was observed with inverted microscope. Apoptosis was evaluated with AO/EB double staining and flow cytometry (FCM); cell cycle was analyzed with FCM .Western blot method was used to detect caspase-3 and caspase-9 protein expression. Results indicated that TVO has significant inhibitory effects on the growth of A431 cells in a dose dependent relationship, the difference between groups has statistically significant (P < 0.05). TVO group compared with control group, concentrations in cells shrivel and broken phenomenon, cell apoptosis rate increased, and a dose dependent and increased the expression of caspase-3 and caspase-9. The experiment results suggested that TVO could restrain skin squamous carcinoma A431 cells proliferation, and induce its apoptosis. The mechanism may be related to increase the expression of caspase-3 and caspase-9.