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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 inhibitory effect of terpinenol 4 on the proliferation of human lung cancer cell line A-549 in vitro and in vivo and its possible mechanism. Methods The concentration of terpinene 4 alcohols was applied to A-549 cells for 24 h. The inhibition rate and IC50 were measured by MTT assay. The growth curves of different concentrations were plotted by CCK-8 colorimetric analysis. The cell ultrastructure was observed by transmission electron microscope after drug treatment. Microstructure changes; flow cytometry was used to detect changes in cell cycle, apoptosis, and intracellular Ca2+ concentration; nude mice implanted with A-549 cells were divided into control group, low-dose group, and high-dose group and injected intraperitoneally for 10 days. The volume and weight of the transplanted tumors in each group were measured, and the anti-tumor effects of the drugs in vivo were observed. Results The IC50 of A-549 cells at 24 h was 0.067% v/v. The results of CCK-8 showed that the growth curve of the treatment group was significantly flatter than that of the control group and was significantly correlated with the dose; the electron microscope showed that terpinolene 4 The number of autophagic vacuoles in alcohol-treated A549 cells increased significantly. FCM detection cycle showed S phase arrest; detection showed that the apoptosis rate increased significantly, the apoptotic rate showed a concentration-effect relationship; the intracellular Ca2+ fluorescence intensity of A-549 cells in each drug intervention group was significantly higher than the control group (P<0.05) . In vivo experiments showed that transplanted tumors grew slowly after treatment with terpinenol in each experimental group. At the end of treatment, the tumor mass and tumor volume in the treated group were significantly lower than those in the control group (P<0.05). The tumor quality was inhibited in the high and low dose groups. The rates were 77.6% and 53.3%, respectively. Conclusion Terpinenol 4 can inhibit the proliferation of A-549 cells in vitro.