松油烯4醇体内外对人肺癌细胞系A-549增殖抑制作用

Effect of Terpinen-4-ol on the proliferation of lung adenocarcinoma A-549 cells in vitro and in vivo

目的探讨松油烯4醇体内外对人肺癌细胞系A-549增殖抑制作用及其可能机制。方法不同浓度松油烯4醇作用于A-549细胞24 h后,应用MTT法检测抑制率及IC50;CCK-8比色分析法绘制不同浓度下的生长曲线;透射电镜观察药物作用后细胞超微结构的变化;流式细胞仪检测细胞周期、凋亡、细胞内Ca2+浓度变化;将A-549细胞移植瘤裸鼠分为对照组、低剂量组、高剂量组药物连续腹腔注射10 d后分别测量各组移植瘤体积和重量,观察药物的体内抗肿瘤作用。结果 A-549细胞24 h的IC50为0.067%v/v;CCK-8实验结果表明给药组生长曲线明显较对照组平缓,并与给药剂量呈显著相关性;电镜观察可见经松油烯4醇处理的A549细胞内自噬泡数量明显增加。FCM检测周期显示S期阻滞;检测显示细胞凋亡率明显增加,凋亡率呈现浓度-效应关系;各药物干预组A-549细胞内Ca2+荧光强度均明显高于对照组(P<0.05)。体内实验显示各实验组经松油烯4醇治疗后移植瘤生长缓慢,治疗结束时治疗组移植瘤质量及瘤体积明显低于对照组(P<0.05),高、低剂量组的瘤质量抑制率分别为77.6%和53.3%。结论松油烯4醇在体外能抑制A-549细胞的增殖。

Objective To investigate the effect of Terpinen-4-ol on the proliferation of lung adenocarcinoma A-549 cells and its related mechanism. Methods A-549 cells were treated with different concentrations of Terpinen-4-ol. The inhibitory effect of Terpinen-4-ol on A-549 cells was tested by MTT method. Cell grow ability was determined by CCK-8 colorimetry. The ultrastructure of A549 cells were observed by transmission electron microscopy before and after Terpinen-4-ol treatment. The changes of cell cycle, apoptosis, and the level of intracel-lular calcium were inspected by flow cytometry. Inoculated the lung adenocarcinoma A-549 cells on the nude mice to form transplantation tumor. The experimental nude mice with transplantation tumors were divided into three groups：negative control group,high dose positive con-trol group and low dose positive control group. The mice were given continuously intraperitoneal injection for 10 days, and then the transplan-tation tumors were taken and the size and weight of them were detected. Results After Terpinen-4-ol treatment for 24 h,MTT assay showed that the IC50 value of A549 cells was 0. 067% v/v. The growth curves of positive control groups were significantly smooth than the negative control group. The formation of autophagosome increased after treatment with Terpinen-4-ol. The results of flow cytometry showed that the cell cycle was arrested in S phase,Terpinen-4-ol could induce apoptosis of A549 cell, The intracellular calcium concentrations in positive control groups were significantly higher than the negative control group（P〈0. 05）. Low dose group and high dose group restrained the growth of the transplantation tumor obviously, and the tumor inhibitory rate were 53. 33% and 77. 76% respectively. Conclusion Terpinen-4-ol has inhibitory effect on the proliferation of A-549 cells in vitro and in vivo.