As_2O_3对人恶性淋巴瘤CA46细胞系增殖、活力和细胞周期的影响及机制探讨  

Effects of Arsenic Trioxide (As_2O_3) on Proliferation,Viability and Cell Life Cycle of Malignant Lymphoma CA46 Cell Line and Related Mechanisms

作　　者：周华蓉[1] 沈建箴[1] 

机构地区：[1]福建医科大学附属协和医院血液科,福州350001

出　　处：《福建医科大学学报》2010年第1期20-23,39,共5页Journal of Fujian Medical University

基　　金：福建省百千万人才工程基金(303052801);福建省自然科学基金(CO540014);福建医科大学教授基金(JS06080)

摘　　要：目的探讨三氧化二砷(As2O3)对恶性淋巴瘤CA46细胞株增殖、活力和细胞周期的影响及机制探讨。方法磺酰罗丹明B(SRB)法观察不同浓度(0.625,1.25,2.5,5.0,10.0,20.0μmol/L)As2O3分别处理24,48,72,96 h后CA46细胞系生长曲线,并计算上述各浓度As2O3处理72 h后CA46细胞系的细胞增殖抑制率;观察不同浓度(0.5,1.0,2.0μmol/L)As2O3作用72 h后CA46细胞系的克隆形成率;流式细胞仪DNA倍体分析法探讨不同浓度(0.5,1.0和2.0μmol/L)As2O3作用72 h对CA46细胞系细胞周期的影响;RT-PCR法检测不同浓度(0.5,1.0和2.0μmol/L)As2O3作用72 h后CA46细胞系p16基因mRNA的表达。同时设相应对照组进行比较。结果(1)与未处理组相比,各浓度As2O3均可明显抑制CA46细胞生长,且G0～G1期细胞逐渐增加,呈剂量依赖性。(2)未处理组细胞p16基因呈微弱表达,不同浓度As2O3作用72 h后,未处理组、不同浓度(0.5,1.0和2.0μmol/L)As2O3处理组p16基因表达阳性条带灰度值与actin比值分别为(0.11±0.11),(0.33±0.10),(0.57±0.11)和(0.67±0.09),各As2O3处理组p16基因mRNA表达明显高于未处理组,其差别有统计学意义(P<0.01)。结论As2O3可将细胞周期阻滞于G0～G1期,抑制肿瘤细胞的增长,其机制可能与诱导p16基因表达有关。Objective To investigate the effects of arsenic trioxide （As2O3） on tb.e ceil protiferation, viability and cell life cycle of malignant lymphoma CA46 cell line and the possible mechanisms. Methods The sulforhodamine B （SRB） method was used to evaluate the growth curve of CA46 cells after the treatment with different concentration（0. 625,1.25,2.5,5.0,10.0,20.0μmol/L） of arsenic trioxide for 24,48,72,96 h, it also was used to calculate the proliferation inhibition ratio of CA46 cells after the treatmen,t with different concentration of As2O3 for 72 hours; to observe the cloning efficiency of CA46 cells after the treatment with different concentration（0. 5,1.0,2. 0μmol/L） of As203 for 72 hours; DNA ploid analytical method was used to analyze the effect of arsenic trioxide on the cell cycle of the target cell line after the treatment with As2O3 （0.5, 1.0 and 2.0 μmol/L） for 72 h; RT-PCR was employed to analyze the expression of p16. Results （1） In comparison with the control group, all 3 different concentrations of arsenic trioxide were able to inhibit the growth of malignant cell line and increase the cell number in G0 or G1 phase in a dose-dependent manner. （2） The expression of p16 gene in the untreated group was weak while that of the treated groups were greatly strengthened after the treatment with As20a for 72 h. The gray scale ratio of p16 to actin treated with As2O3 （0. 5, 1.0 and 2.0 μmol/L） was increased respectively from （0.11±0.11） to （0.33±0.10）, （0.57±0.11）, （0.67±0.09）, exhibiting a significant difference （P〈0. 01）. Conclusion Arsenic trioxide may achieve cell cycle control and cell growth retardation in malignant cells through the demethylation of p16, which finally blocks the cells at G0 or G1 phase.

关 键 词：砷剂 淋巴瘤 细胞增殖 肿瘤细胞 培养的 细胞周期 基因 P16 

分 类 号：R73-3[医药卫生—肿瘤]