长春瑞滨诱导人肺癌Calu-3细胞凋亡及机制

Vinorelbine-induced apoptosis of cultured human lung cancer cells and mechanisms

目的 观察长春瑞滨 (VRB)诱导人肺癌Calu 3细胞凋亡时Bcl 2和半胱天冬酶 3的表达有无变化。方法 以不同浓度的VRB( 2 0 ,40和 60 μmol·L-1)作用于体外培养的人肺癌Calu 3细胞 2 4h后 ,TUNEL法和吖啶橙染色法观察肺癌细胞凋亡形态学特征 ;流式细胞仪检测肺癌细胞凋亡率和肺癌细胞Bcl 2蛋白表达水平 ;以半胱天冬酶 3荧光分析检测试剂盒测定肺癌细胞半胱天冬酶 3活性。结果 VRB( 2 0 ,40和60 μmol·L-1)处理细胞 2 4h ,TUNEL法及吖啶橙染色均观察到典型的凋亡细胞形态学特征。流式细胞仪检测VRB处理的肺癌细胞凋亡率分别为 ( 3 .1±0 .6) % ,( 7.8± 1 .2 ) %和( 1 9.6± 4.3 ) % ,较对照组 (凋亡率为 0 )显著增高且呈剂量依赖性 (P <0 .0 1 ) ;Bcl 2蛋白阳性表达细胞率分别为 ( 3 7.6±6.9) % ,( 2 5 .4±6.2 ) %和( 8.4±2 .5 ) % ,较对照组 ( 4 8.3±7.1 ) %显著降低且呈剂量依赖性 (P <0 .0 5 ) ;肺癌细胞半胱天冬酶 3活性分别为 ( 3 3 2± 1 6) ,( 4 1 7± 1 1 )和 ( 63 1± 2 7)μmol·L-1·h-1,较对照组 ( 1 95±1 2 ) μmol·L-1·h-1显著增高且呈剂量依赖性(P <0 .0 1 )。结论 VRB可以诱导肺癌细胞凋亡 ,抑制Bcl 2表达及增强半胱天冬酶

AIM To study if there were any changes of Bcl 2 expression and caspase 3 activity in vinorelbine(VRB) induced apoptosis of human lung cancer cell line Calu 3. METHODS Cells were incubated with VRB(20, 40 and 60 μmol·L -1 ) for 24 h. Morphological changes in apopto tic cells were studied by TUNEL FITC staining and acridine orange(AO) staining. Flow cytometer was used to detect apoptotic rates and Bcl 2 expression. Caspase 3 activity was detected by spectrofluorometer. RESULTS After treatment with VRB (20, 40 and 60 μmol·L -1 ) for 24 h, typical morphological features of apoptotic cells were appeared. Apoptotic rates of different concentrations of VRB treated cells were (3.1± 0.6) %,(7.8±1.2)% and (19.6±4.3)%, respectively, significantly higher than that of control cells(0%, P<0.01). Bcl 2 expression rates of VRB treated cells were (37.6±6.9)%, (25.4±6.2)% and (8.4±2.5)%, respectively, much lower than that of control ones(48.3±7.1)% (P<0.05, P<0.01). Caspase 3 activity of VRB treated cells were (332±16), (416±11) and (631±27)μmol·L -1 ·h -1 , respectively, significantly higher than that of control ones (195±12) μmol·L -1 ·h -1 (P<0.01). All changes were in a concentration dependent manner. CONCLUSION VRB can induce apoptosis of human lung cancer cells Calu 3 effectively through inhibiting Bcl 2 protein and activation of caspase 3 activity.