摘要
目的 :研究拓扑特肯 (Topotecan ,TPT)诱导体外培养的人肺癌SPC A 1细胞凋亡的作用及其可能的机制。方法 :体外培养肺癌细胞 ,加以 2 0mg/L的TPT孵育 12、2 4和 4 8h ,用MTT法检测TPT对肺癌细胞增殖的作用 ;用TUNEL法和流式细胞仪技术检测TPT诱导肺癌细胞凋亡的发生情况及Cas pase 3特异性拮抗剂Ac DEVD CHO (5 0 μmol/L)对TPT诱导凋亡作用的影响。 结果 :TPT处理细胞12、2 4和 4 8h后 ,肺癌细胞的存活率分别为 (85 8± 9 7) %、(76 5± 6 5 ) %和 (6 4 6± 10 4 ) % ,未经TPT处理的对照组细胞存活率为 (97 1± 6 9) % ,各组间差异有显著性 (P <0 0 5 ) ;流式细胞仪检测其凋亡率分别为 (3 4± 1 5 ) %、(11 5± 2 8) %和 (31 2± 4 1) % ,各组间差异有显著性 (P <0 0 1) ;同时TUNEL法可检测到典型的肺癌细胞凋亡形态学特征 ;Ac EDVD CHO和TPT共孵育细胞 12、2 4和 4 8h后其凋亡率分别为 0、(5 .5± 1.6 ) %和 (18.6± 3.7) % ,较对应的未加Ac DEVD CHO组明显降低 (P <0 .0 1)。结论 :TPT可通过Caspase 3依赖的途径诱导体外培养的人SPC A 1肺癌细胞凋亡 ,且与作用时间呈正相关。
Objective To study the Topotecan(TPT) induced apoptosis of human SPC A 1 lung cancer cells and its possible mechanism.Methods Cells were incubated with TPT(20 mg/L) for 12,24 and 48 h.Cell viability was measured by MTT assay. Morphological changes of apoptotic cells was studied by TUNEL staining. Apoptotic rates of cells incubated with TPT or TPT and 50 μmol/L Ac DEVD CHO(specific Caspase 3 inhibitor)were detected by flow cytometer.Results Cell viability with 20 mg/L TPT for 12,24 and 48 h were (85.8±9.7)%, (76.5±6.5)% and (64.6±10.4)% respectively.Cell viability without TPT was (97.1±6.9)%.There was a significant reduction of cell viability with time duration of TPT( P <0 05).Typical morphological features of apoptotic cells were detected by TUNEL staining.Apoptotic rates of cells with TPT for 12,24 and 48 h studied by flow cytometer were (3.4±1.5)%,(11.5±2.8)% and (31.2±4.1)% respectively. There was a significant rising apoptotic rates with time duration( P <0 01). Apoptotic rates of cells with TPT and Ac DEVD CHO for 12,24 and 48 h were 0,( 5.5±1.6)% and (18.6±3.7)% respectively,lower than cells without Ac DEVD CHO( P <0 01).Conclusions TPT can act effectively as chemotherapeutic drugs for lung cancer by leading cancer cells to apoptosis through Caspase 3 activation and the function is time dependent.
出处
《肿瘤防治杂志》
2002年第3期249-251,共3页
China Journal of Cancer Prevention and Treatment
