卵巢癌细胞系顺铂化疗敏感株与耐药株中ZNF217的表达

Expressions of zinc-finger protein 217(ZNF217)in DDP-sensitive and-resistant human ovarian cancer cell lines

目的探讨锌指蛋白(ZNF)217在卵巢癌细胞系顺铂敏感株与耐药株中表达的差异。方法选择6株卵巢癌细胞,其中A2780、SKOV-3和COC1为顺铂敏感株,A2780-DDP-R、SKOV-3-DDP-R和COC1-DDP-R为顺铂耐药株。ATP法检测细胞的相对发光单位(RLU),计算顺铂对细胞的半效抑制浓度(IC50)和各耐药株的耐药指数(RI)。免疫荧光细胞化学法观察ZNF217在细胞内的定位,RT-PCR和Westernblotting法检测ZNF217mRNA和蛋白的表达水平。结果顺铂对A2780和A2780-DDP-R的IC50分别为18.1±2.3mg/L和47.9±3.8mg/L(P<0.01),对SKOV-3和SKOV-3-DDP-R为9.6±1.5mg/L和40.1±5.3mg/L(P<0.01),对COC1和COC1-DDP-R为4.8±0.9mg/L和15.3±2.8mg/L(P<0.01);A2780-DDP-R、SKOV-3-DDP-R、COC1-DDP-R对顺铂的RI分别为2.6、4.2和3.2。激光共聚焦显微镜可见,ZNF217蛋白在敏感株中主要分布于细胞质,在耐药株则主要分布于细胞核。耐药株中ZNF217mRNA和蛋白表达均明显高于敏感株。结论ZNF217的高表达和核转位可能与卵巢癌对顺铂耐药有关。

Objective To explore the mRNA and protein expressions of zinc-finger protein 217 （ZNF217） in cisplatin （DDP）- sensitive and DDP-resistant human ovarian cancer cell lines. Methods Six strains of ovarian cancer cells, including 3 DDP-sensitive strains （A2780, SKOV-3 and COC1） and 3 DDP resistant strains （A2780-DDP R, SKOV-3-DDP-R and COC1-DDP R） were selected. The relative luminescence unit （RLU） of cells was determined with ATP assay, the IC50 value and resistance index （RI） of DDP resistant cell lines were calculated. Intracellular localization of ZNF217 protein in the 6 ovarian tumor cell lines was detected by immunofluoreseent cytochemistry. The expressions of ZNF217 mRNA and protein were determined by RT-PCR and Western blotting, respectively. Results The IC50 values of DDP to A2780 and A2780-DDP R were 18. 1±2. 3mg/L and 47. 9±3. 8rng/L （P〈0. 01）, respectively, that m SKOV- 3 and SKOV 3-DDP R were 9. 6 ± 1.5mg/L and 40. 1 ± 5. 3mg/L （ P〈0. 01）, respectively, and that to COC1 and COCl-DDP-R were 4. 8±0. 9mg/L and 15. 3± 2.8mg/L （ P〈0. 01 ）, respectively. The RI of A278（〉DDP R, SKOV 3-DDP R and COC1-DDP-R to DDP were 2. 6, 4. 2 and 3. 2, respectively. Laser-scanning confocal microscopy showed that ZNF217 protein was mainly located in the cytoplasm of the DDP-sensitive cell lines, but it was in the nuclei of the DDP-resistant cell lines. RT-PCR and Western blotting showed that the expressions of both ZNF217 mRNA and protein were significantly higher in DDP-resistant cell lines than in DDP-sensitive cell lines. Conclusion Over-expression and nuclear translocation of ZNF217 may be related to the resistance of ovarian cancer cells to DDP.