ATP柠檬酸裂解酶表达下调对前列腺癌细胞生长及凋亡影响的研究  

Effects of downregulated ATP citrate lyase on the growth and apoptosis of prostate cancer cells

作　　者：孔海瑞[1] 耿申[2] 杨杰[1] 曾方银[3] 

机构地区：[1]南方医科大学南方医院检验科,广州医学硕士510515 [2]南方医科大学金陵医院(南京军区南京总医院)呼吸内科,南京210002 [3]南方医科大学第五附属医院检验科,广州510900

出　　处：《医学研究生学报》2017年第1期26-30,共5页Journal of Medical Postgraduates

基　　金：广东省自然科学基金(S2013010014482)

摘　　要：目的肿瘤细胞可通过改变代谢模式来支持自身的恶性增殖,前列腺细胞更多地依赖于脂质代谢,而ATP柠檬酸裂解酶(ACLY)在脂质代谢中占有非常重要的位置。文中探讨ACLY表达下调对激素非依赖型人前列腺癌DU145细胞增殖能力及周期分布、凋亡的影响。方法将DU145细胞分为实验组和对照组,实验组使用小干扰RNA片段选择性敲低ACLY的表达,对照组加入无意义小干扰RNA片段,采用细胞增殖与活性检测技术CCK-8观察ACLY分子表达下调对DU145细胞增殖能力的影响,流式细胞仪分析不同处理组间细胞周期分布及凋亡细胞比例之间的的差异,Western blot法检测胞内Caspase-3蛋白含量的变化。结果 Western blot结果显示ACLY干扰效果良好,与对照组相比,实验组细胞内ACLY蛋白含量下降明显(P<0.05)。与对照组相比,实验组细胞内乙酰辅酶A含量、早期凋亡及晚期凋亡细胞所占的比例明显升高[(0.42±1.99)vs(0.79±2.38),(37.10±3.28)vs(6.20±2.88),P<0.05]。在DU145细胞内选择性敲低ACLY的表达含量,实验组较对照组细胞增殖能力减弱,并且随着时间的推移表现更加明显。在48 h及72 h这两个时间点,2组细胞同一时间点吸光度差异均具有统计学意义(P<0.05)。在DU145细胞内干扰ACLY的表达,实验组与对照组相比,G1期细胞所占比例增高,但差异无统计学意义(P>0.05)。G2期细胞占比降低,S期比例增高,细胞周期阻滞在G0/G1期,两组之间的差异具有统计学意义(P<0.05)。同时实验组细胞内凋亡蛋白Caspase-3的表达明显上调。结论 ACLY对维持前列腺癌细胞恶性增殖具有重要的意义,其表达下调能抑制DU145细胞的分裂增殖并促进细胞的凋亡。Objective Tumor cells are able to support their malignant proliferation by changing metabolic models. Prostate cells rely much on lipid metabolism in which ATP-citrate lyase（ ACLY） plays a very important role. The aim of this research was to study the effects of downregulated ACLY on the cell proliferation,cycle distribution and apoptosis of androgen-independent prostate cancer cells DUl45. Methods DU145 cells were divided into two groups： the cells in experiment group were transfected with the small interfering RNA-mediated knockdown of ACLY,while the cells in control group were transfected with meaningless small interfering RNA. Cell counting Kit test（ CCK-8） was applied to detect the effects of the downregulation of ATP citrate lyase on the proliferation of DU145. Flow cytometry instrument was used to analyze the variation of cell cycle distribution and apoptosis rate between groups. Western blot was used to detect the change of intracellular Caspase- 3 protein content. Results Western blot showed favorable effects of ACLY interference. Compared with control group,ACLY protein content significantly decreased in experiment group（ P〈0.05）; the acetyl coenzyme A content and the percentage of early apoptosis cells and late apoptosis cells increased significantly [（ 0.42±1.99） vs（ 0.79±2.38）,（ 37.10±3.28） vs（ 6.20±2.88）,P〈0.05]. As to the selective ACLY knockdown in DU145 cells,the proliferation ability in experiment group weakened in comparison with control group,which became more and more apparent as time went on. At 48 h and 72 h time points,the cell absorbance between two groups at the same time point was of significant difference（ P〈0.05）. As to the interfering ACLY expression in DU145 cells,the percentage of G1 cells increased without any statistical significance（ P〉0.05）,while the percentage of G2 cells decreased and the percentage of S cells increased with most cell cycle blocking at G0/G1 stage,which were of significant difference. Meanwhile the expression

关 键 词：前列腺癌 ATP柠檬酸裂解酶 凋亡 增殖 细胞周期 

分 类 号：R737.25[医药卫生—肿瘤]