ZHOU Ding-an,HUANG Wen-fang,ZHANG Ming-chuan,ZHAO Shen,XU Yi.Molecular changes of PI3K-AKT signal pathway in K562 cells treated with simvastatin in vitro and in vivo[J].J Amry Med Univ (J Third Mil Med Univ),2007,29(21):2049-2055.
ZHOU Ding-an,HUANG Wen-fang,ZHANG Ming-chuan,ZHAO Shen,XU Yi.Molecular changes of PI3K-AKT signal pathway in K562 cells treated with simvastatin in vitro and in vivo[J].J Amry Med Univ (J Third Mil Med Univ),2007,29(21):2049-2055.DOI:
Molecular changes of PI3K-AKT signal pathway in K562 cells treated with simvastatin in vitro and in vivo
K-AKT信号通路的分子变化,以分析辛伐他汀诱导K562细胞凋亡的机制。 方法 体外培养CML 细胞株K562细胞,MTT噻唑蓝法检测细胞增殖抑制率。流式细胞术检测辛伐他汀作用后的细胞周期和凋亡率的变化。另外,12只BALB/c-nu/nu裸小鼠皮下接种K562细胞,构建Balb/c-nu/nu裸小鼠的慢性粒细胞白血病动物模型,采用缺口末端标记技术TUNEL(TdT-mediated dUTP nick end labeling)法检测辛伐他汀诱导K562细胞早期凋亡的变化,用RT-PCR检测辛伐他汀在体内外K562细胞后N-ras分子和N-ras分子下游的PI
K-AKT-dependent manner. Methods K562 cells were cultured in vitro for experiments below: the proliferation assay of K562 cells detected by MTT
the analysis of apoptosis rate and cell cycle of K562 cells measured by flow cytometry (FCM)
the construction of chronic myeloid leukemia (CML) animal model through the subcutaneous inoculation of K562 cells to 12 BALB/c-nu/nu nude mice
the early apoptosis changes of K562 cells detected by TdT-mediated dUTP nick end labeling (TUNEL) and the in vitro and in vivo changes of N-ras
PI
3
K
AKT1
IKK-β
NF-κB1 at transcription level detected by RT-PCR. Results Simvastatin inhibited the proliferation of K562 cells and induced their G
0
/G
1
arrest and significant apoptosis. N-ras and most genes of PI
3
K-AKT signal pathway were expressed differentially in vitro. K562 cells on nude mice could be induced to apoptosis by simvastatin and the apoptotic index increased with the dose accumulation of simvastatin (P=0.000). The differential mRNA expression changes of N-Ras and most genes of PI
3
K-AKT signal pathway in K562 cells were observed after treatment of simvastatin at different doses (P=0.000 or P=0.003). However
mRNA expression of the genes in PI
3
K-AKT pathway in vitro differed to that in vivo. Conclusion The differential expression at transcription levels of N-ras and the most genes in PI
3
K-AKT pathway that are involved in anti-apoptosis in K562 cells
to some degree
indicates that simvastatin can induce the apoptosis of K562 cells in a PI3K-AKT pathway-dependent fashion in vitro. The animal experiments confirm that there are different mechanisms of simvastatin in inducing the apoptosis of K562 cells in vitro and in vivo.