TRAIL联合依托泊甙对骨肉瘤OS732细胞增殖、凋亡的影响

摘要

目的 观察TNF相关凋亡诱导配体(TRAIL)联合依托泊甙对骨肉瘤OS732细胞增殖、凋亡的影响.方法 对数生长期的OS732细胞接种于96孔培养板中培养,待细胞完全贴壁后给药.将细胞分为三组,TRAIL组分别加10、50、100、1 000 ng/mL的TRAIL；依托泊甙组分别加1、5、10、20 μg/mL的依托泊甙,协同组同时加50ng/mL 的TRAIL与5μg/mL的依托泊甙.采用MTT法检测各组细胞增殖抑制率,倒置显微镜下观察OS732细胞形态学变化,荧光显微镜下观察细胞凋亡情况.结果 TRAIL组在10、50、100、1 000 ng/mL TRAIL作用下OS732细胞增殖抑制率分别为6.47％ ±0.76％、9.85％ ±0.97％、15.24％ ±2.48％、51.32％ ±4.71％(P均＜0.05),依托泊甙组在1、5、10、20 μg/mL依托泊甙作用下OS732细胞增殖抑制率分别为3.67％±0.74％、10.15％ ±2.62％、29.37％±3.36％、43.59％±5.92％(P均＜0.05).协同组细胞增殖抑制率为48.36％±6.42％,高于单用TRAIL( 50ng/mL)时的9.85％±0.97％及单用依托泊甙(5 μg/mL)时的10.15％±2.62％(P均＜0.05).倒置相差显微镜下见协同组大部分细胞脱落,悬浮于培养液中,少部分细胞破裂呈坏死状,荧光显微镜下见协同组细胞内发亮的荧光显色及荧光浓聚,提示有大量OS732细胞发生凋亡.结论 TRAIL、依托泊甙联用可抑制骨肉瘤OS732细胞增殖,促进其凋亡,效果优于其单药应用.%Objective To study the inhibition effect of combination of TRAIL and etoposide on the osteosarcoma cell lines and explore the nerw therapy of the osteosarcoma cell 732 ( OS732). Methods Logarithmic phase OS732 cells were seeded in 96-well culture plates in culture. The cells were divided into three groups, TRAIL group was treated with TRAIL of 10, 30, 100 and 1 000 ng/mL; etoposide group was treated with etoposide of 1, 5, 10, 20 (jg/mL; collaborative group was treated with 50 ng/mL TRAIL and 5 ^tg/mL etoposide. MTT assay was used to detect the cell proliferation inhibition rate, morphological changes and cell apoptosis were observed. Results Inhibition rate of cell proliferation of OS732 in TRAIL group treated with 10, 50, 100, 1 000 ng/mL of TRAIL were 6.47% ±0.76%, 9.85% ±0.97%, 15.24% ± 2.48% , 51.32% ±4.71% , respectively (all P<0.05). Inhibition rate of cell proliferation of OS732 in etoposide group treated with 1, 5, 10, 20 jig/mL etoposide were 3.67% ±0.74%, 10.15% ±2.62%, 29.37% ±3.36%, 43.59% ± 5.92% , respectively( all P < 0.05 ). Inhibition rate of cell proliferation of OS732 in collaborative group was 48. 36% ± 6.42% , higher than that of 9.85% ±0.97% with TRAIL (50 ng/mL) alone, and 10.15% ±2.62% with etoposide(5 )ig/mL) alone (all P <0.05). Most of the cells in collaborative group were found off suspended in culture medium, and small number of cell rupture were necrosis. Conclusion Combination of TRAIL and etoposide can inhibit the cell proliferation of OS732, and promote apoptosis, the effect is better than single drug application.