摘要
目的探讨石蒜碱对肾癌细胞ACHN的作用及可能机制。方法流式细胞仪分析石蒜碱对ACHN细胞周期及凋亡的影响;MTS检测石蒜碱对ACHN细胞增殖的作用;划痕实验、Transwell实验检测石蒜碱对ACHN细胞迁移、侵袭能力的影响;平板克隆形成检测石蒜碱对ACHN细胞集落形成能力的影响;q RT-PCR分析基因转录的变化;Western blotting分析蛋白表达水平的变化。结果石蒜碱对ACHN半数抑制浓度为24.34μmol/L。石蒜碱能将肾癌ACHN细胞剂量依赖性阻滞于G0/G1期(P<0.01);与正常对照组相比,5μmol/L石蒜碱即能诱导ACHN细胞凋亡(P<0.01)、抑制ACHN细胞增殖(P<0.01);处理后ACHN细胞迁移能力(P<0.01)、侵袭能力(P<0.01)显著下降、集落形成能力亦呈剂量依赖性降低。q RT-PCR及Western blotting显示5μmol/L的石蒜碱即能在基因及蛋白水平上调CDK4、Bax表达,而下调Cyclin D1、Bcl-2、Survivin的表达;caspase-3基因表达增加,caspase-3蛋白水平变化却不甚明显。结论石蒜碱对肾癌ACHN细胞具有明显的抗肿瘤效应,该药可为肾癌的治疗提供新思路。
Objective To investigate the antitumor effect of lycorine on renal cell carcinoma ACHN cells and explore the possible mechanism. Methods We used flow cytometry to examine the effect of lycorine on ACHN cell cycle and apoptosis. The cell proliferation, migration and invasion were assessed with MTS assay, wound healing assay, and Transwell assay, respectively.Colony forming assay was performed, and the m RNA and protein levels of Bax, Bcl- 2, survivin, caspase- 3, cyclin D1 and CDK4 were measured with q RT- PCR and Western blotting. Results Lycorine obviously inhibited the proliferation of ACHN cells with an IC50 of 24.34 μmol/L. Lycorine also induced apoptosis of ACHN cells, caused cell cycle arrest at G0/G1 phase, and suppressed the colony forming ability of the cells in a dose- dependent manner. The migration and invasion of ACHN cells were significantly inhibited by 5 μmol/L lycorine. Lycorine up- regulated the m RNA levels of CDK4, Bax, caspase- 3 while down-regulated the levels of survivin, Bcl-2 and Cyclin D1; the protein levels of CDK4 and Bax were increased and cyclin D1,Bcl- 2 and surviving expressions were decreased, but caspase- 3 expression showed no significant changes following the treatment. Conclusion Lycorine has obvious antitumor effect against ACHN cells, suggesting its value as a new therapeutic agent for renal cell carcinoma.
出处
《南方医科大学学报》
CAS
CSCD
北大核心
2016年第6期857-862,共6页
Journal of Southern Medical University
基金
广州市珠江科技新星项目(2013J2200044)
关键词
石蒜碱
肾癌
凋亡
细胞周期
renal cell carcinoma
lycorine
apotosis
cell cycle
