胡桃醌通过降低AKT活性抑制肺癌A549细胞增殖  

The inhibition effect of Juglone on proliferation of lung cancer A549 cells through down-regulation of AKT activity

作　　者：刘元银[1] 雷淑慧 杨燕[3] 蒋幼凡[1] 

机构地区：[1]重庆医科大学附属第二医院呼吸内科,重庆400010 [2]铜梁县人民医院重症监护科,重庆402560 [3]攀枝花市中心医院呼吸内科,四川攀枝花617067

出　　处：《基础医学与临床》2013年第8期1032-1037,共6页Basic and Clinical Medicine

摘　　要：目的探讨肽酰脯氨酰顺反异构酶(PIN1)抑制剂胡桃醌(Juglone)对肺癌A549细胞增殖的影响及分子机制。方法用胡桃醌(0、6.25、12.5、25和50μmol/L)处理A549细胞,MTT法检测细胞增殖,流式细胞术检测细胞凋亡及周期,real-time PCR检测PIN1、AKT mRNA表达,Western blot检测PIN1、AKT及AKT-pS473蛋白表达。结果胡桃醌能浓度依赖性抑制A549细胞增殖,促进细胞凋亡,将细胞周期阻滞在G2/M期和S期。与对照组相比,各处理组PIN1 mRNA水平随着胡桃醌浓度增加而降低(P<0.05)。不同浓度胡桃醌处理组PIN1蛋白表达为1.032±0.056、0.892±0.024、0.596±0.023和0.396±0.021,均显著低于对照组的1.280±0.046(P<0.05);AKT-pS473蛋白表达为0.554±0.023、0.464±0.018、0.362±0.015和0.228±0.020,均显著低于对照组的0.626±0.015(P<0.05);48 h处理组PIN1及AKT-pS473蛋白表达分别为0.575±0.036和0.338±0.014,显著低于24 h的0.764±0.032和0.436±0.023(P<0.05)。结论 PIN1抑制剂胡桃醌可以抑制肺癌A549细胞的增殖,其机制可能是通过降低AKT-pS473活性来实现的。Objective To explore the effect and mechanism of PIN1 inhibitor Juglone on cell proliferation of human lung cancer cells A549 in vitro. Methods 21549 cells were treated by different concentrations of Juglone. Cell pro- liferation was measured by MTT assay respectively. The apoptosis rate and cell cycle of A549 cells were detected by flow cytometry. Real-time PCR was carried to detect the mRNA expression of PIN1 and AKT. Western blot were used to observe the protein expression of PIN1 and AKT-pS473 in cells. Results With the increasing of Juglone＇s concentration, the inhibition effect on proliferation and the apoptosis rate of A549 cells enhanced in a dose-depen- ded manner, and A549 cells cycle were blocked in G2/M and S period. Real-time PCR result showed the mRNA level of PIN1 in A549 cells treated by Juglone significantly decreased compared with control group. Western blot showed that the protein expression of PIN1 in Juglone groups were 1. 032 ± 0. 056, 0. 892 ± 0. 024, 0. 596 ± 0. 023, 0. 396 ± 0. 021, significantly lower than that of control group 1. 280 ± 0. 046 （P 〈 0. 05 ）. the protein ex- pression of AKT-pS473 in Juglone groups was 0. 554 ± 0. 023, 0. 464±0. 018, 0. 362 ± 0. 015, 0. 228 ± 0. 020, significantly lower than control group 0. 626 ± 0. 015 （P 〈 0. 05 ）. the protein expression of PIN1 and AKT-pS473 in cells treated by Juglone for 48 h were 0. 575 ±0. 036 and 0. 338 ±0. 014, significantly lower than 0. 764 ±0. 032 and 0. 436 ± 0. 023 of 24 h group （ P 〈 0. 05 ）. Conclusions Inhibiting the expression of PIN1 with Juglone can significantly depress the proliferation activity of A549 through decreasing the phosphorylation level of AKT.

关 键 词：肽酰脯氨酰顺反异构酶 胡桃醌 细胞增殖 肺癌 

分 类 号：R734.2[医药卫生—肿瘤]