【摘要】 目的　探讨凋亡相关基因和半胱天冬氨酸蛋白酶 3 (caspase 3 )在卵巢癌顺铂 (DDP)耐药细胞株中的表达。方法　采用逆转录聚合酶链反应 (RT PCR)技术、蛋白印迹法和流式细胞仪分析卵巢癌DDP敏感细胞株A2 780、COC1和DDP耐药细胞株A2 780 /DDP、COC1/DDP中凋亡相关基因bcl 2、bcl XL、bax和bcl Xs的表达 ,及不同浓度DDP(0、5、10、2 0 μmol/L)作用后上述 4种细胞中caspase 3活性及其底物多聚腺苷二磷酸核糖聚合酶 (PARP)的变化。结果　bcl 2和bcl XLmRNA的表达 ,A2 780 /DDP细胞分别为 1 87± 0 2 5和 1 73± 0 15 ,明显高于A2 780细胞的 1 48± 0 14和 1 41± 0 19,差异有显著性 (P <0 0 5 ) ;bcl 2和bcl XL 蛋白的表达 ,A2 780 /DDP细胞分别为 1 99± 0 11和 1 69± 0 16,明显高于A2 780细胞的 1 5 1± 0 17和 1 2 8± 0 11,差异有显著性 (P <0 0 5 ) ;bcl 2和bcl XL在COC1/DDP细胞中的表达明显高于COC1细胞 ,差异有显著性 (P <0 0 5 ) ;bax的表达 ,A2 780 /DDP与A2 780、COC1/DDP与COC1细胞分别比较 ,差异均无显著性 (P >0 0 5 ) ;在上述 4种细胞中均未检测到bcl Xs的表达。不同浓度DDP作用后 ,A2 780 /DDP和COC1/DDP细胞中caspase 3活性明显低于A2 780和COC1细胞 (P <0 0 5 ) ;其凋亡率更多还原

【Abstract】 Objective To explore the role of apoptosis associated genes and caspase 3 activity in cisplatin(DDP) resistant human ovarian cancer cell lines Methods The expressions of apoptosis associated genes(bcl 2, bax, bcl X L and bcl X S)and the activity of caspase 3 were studied by reverse transcription polymerase chain reaction (RT PCR) and western blot in the cisplatin resistant (A2780/DDP, COC1/DDP) and sensitive human ovarian cancer cells(A2780 and COC1) The apoptotic rates of A2780, COC1, A2780/DDP and COC1/DDP were measured with flow cytometry when treated with cisplatin Results The mRNA expressions of bcl 2 and bcl X L in A2780/DDP cells were 1.87±0.25 and 1.73±0.15, and significantly higher than those in A2780 cells ( P <0.05), which were 1.48±0.14 and 1.41±0.19 respectively. The protein expressions of bcl 2 and bcl X L in A2780/DDP cells were 1.99±0.11 and 1.69±0.16, and significantly higher than those in A2780 cells ( P <0.05), which were 1.51±0.17 and 1.28±0.11 respectively. The expressions of bcl 2 and bcl X L in COC1/DDP cells were also significantly higher than those in COC1 cells ( P <0.05). While there was no significant difference in expression of bax between A2780/DDP and A2780 as well as between COC1/DDP and COC1. No expression of bcl X S was detected. When cells treated with different concentration of DDP, the caspase 3 activities, apoptotic rates and PARP expressions in A2780/DDP and COC1/DDP were significantly lower than those in A2780 and COC1 ( P <0.05), which showed dose dependent ( P <0.05).Conclusion Overexpression of anti apoptotic genes and decrease of caspase 3 activity may relate to cisplatin resistance in human ovarian cancer cell lines更多还原