榄香烯联合热疗对人肺腺癌A549细胞增殖及p-Akt表达的影响

摘要

目的 探讨榄香烯(ELE)联合热疗(HTM)对人肺腺癌A549细胞形态、增殖、细胞周期及p-Akt蛋白表达的影响.方法 采用倒置光学显微镜观察ELE(40μg/ml)与HTM单独或联合对人肺腺癌A549细胞形态学的影响；MTT法检测ELE(20、40、80、160μg/ml)与HTM单独或联合对人肺腺癌A549细胞增殖的抑制作用；流式细胞术检测ELE(40μg/ml)与HTM单独或联合对人肺腺癌A549细胞周期的影响；Western blotting检测ELE(40μg/ml)与HTM单独或联合作用于人肺腺癌A549细胞24h后p-Akt蛋白的表达.结果 倒置光学显微镜结果显示,HTM组、ELE组和ELE联合HTM组的细胞数均明显减少,部分细胞变圆、体积变小、核固缩,但ELE联合HTM组变化最明显,活细胞数最少.与HTM组和ELE组比较,ELE联合HTM对A549细胞增殖的抑制作用显著增强(P＜0.05),呈时间和剂量依赖性；ELE联合HTM作用于人肺腺癌A549细胞24、48和72h的IC50分别为88、65和37μg/ml,低于单独ELE的103、81和59μg/ml.ELE联合HTM组作用24h后肺腺癌A549细胞S期的比例为(52.07 ±3.10)％,显著高于HTM组的(33.40±0.87)％和ELE组的(41.58±3.21)％ (P ＜0.05).ELE联合HTM组p-Akt蛋白的相对表达量为0.52±0.01,显著低于HTM组的0.99 ±0.04和ELE组的0.69±0.03(P ＜0.05).结论 ELE联合HTM能增强对人肺腺癌A549细胞的增殖抑制作用,下调p-Akt蛋白表达可能是其作用机制之一.%Objective To investigate the effects of elemene (ELE) combined with hyperthermia (HTM) on the morphology, proliferation, cell cycle and p-Akt expression of human lung adenocarcinoma A549 cells. Methods Cytomorphology of human lung adenocarcinoma A549 cells treated by ELE(40μg/ml) or HTM alone or in combination was observed by inverted optical microscope. The proliferation of human lung adenocarcinoma A549 cells treated by ELE(20, 40, 80, 160μg/ml) or HTM alone or in combination was assessed by MTT. Cell cycles of human lung adenocarcinoma A549 cells treated by ELE or HTM alone or in combination were detected by flow cytometer. The expression of p-Akt treated by ELE or HTM alone or in combination was determined by Western blotting. Results Under optical microscope, the number of tumor cells in the ELE, HTM and ELE combined with HTM groups decreased obviously, and some of them became round and smaller with nucleus pyknosis, especially in the combination group. Compared with sole ELE and HTM, ELE combined with HTM group showed stronger inhibition to the proliferation of human lung adenocarcinoma A549 cells in a time-dose dependent manner( P < 0. 05). The IC50 in combination group was 88, 65 and 37μg/ml at 24, 48 and 72h, lower than 103, 81 and 59μg/ml of sole ELE group. Tumor cells of (52. 07 ±3. 10)% were arrested at the S-phase in combination group, higher than(33. 40 ±0. 87) % in solo HTM group and(41. 58 ±3.21) % in ELE group(P <0. 05). The p-Akt expression was 0. 52 ± 0. 01, lower than 0. 99 ± 0. 04 in HTM group and 0. 69 ± 0. 03 in ELE group (P < 0. 05). Conclusion ELE combined with HTM enhances the inhibition to the proliferation of human lung adenocarcinoma A549 cells, and the mechanism may be related to the down-reg-ulatin of p-Akt.