谷胱甘肽转移酶π基因在COC1/DDP细胞对顺铂耐药的作用  

Effect of glutathione S-transferase π in COC1/Cisplatin cells on cisplatin resistance

作　　者：张珂[1] 杨兴升[1] 孙彩萍[1] 胡滨[1] 

机构地区：[1]郑州大学第二附属医院妇产科,450002

出　　处：《中华实验外科杂志》2015年第4期874-876,共3页Chinese Journal of Experimental Surgery

摘　　要：目的 观察顺铂耐药卵巢癌细胞株COC1/DDP中谷胱甘肽转移酶π（GSTπ）基因的表达,探讨其反义寡核苷酸对COC1/DDP顺铂耐药的逆转作用.方法 设计合成针对GSTπ的硫代脱氧寡核苷酸（ASODN）,以200 nmol/L浓度转染COC1/DDP细胞,用噻唑蓝法观察顺铂对细胞抑制率的变化,反转录-聚合酶链反应（RT-PCR）检测基因在mRNA水平表达的改变.结果 （1）COC1/DDP细胞中GSTπ基因呈高表达,其mRNA相对表达值为0.78±0.13.而COC1细胞中无表达.（2）与COC1/DDP细胞组比较,GSTπ-ASODN组GSTπ-mRNA降低,相对表达值为0.058±0.022,耐药指数（RI）降低为3.61,差异均有统计学意义（P＜0.05）;错义寡核苷酸组,GSTπ表达及RI改变差异无统计学意义（P＞0.05）.结论 （1）GSTπ基因高表达与COC1细胞获得顺铂耐药性有密切关系.（2） GST-ASODN能有效逆转COC1/DDP细胞株对顺铂的耐药性.Objective To study the expression of drug resistance related gene in COC1/Cisplatin （DDP） cell line,and the reversion of cisplatin resistance by antisense oligodeoxynucleotide （ASODN） of the overexpressed gene.Methods The expression of glutathione S-transferase π （GSTπ） in COC1/DDP cell line was examined by reverse transcription-polymerase chain reaction （RT-PCR）,and compared with that in COC1 cell line.ASODN targeting GSTπ was designed and synthesized.GSTπ-ASODN and missense oligodeoxynucleotide with the concentration of 200 nmol/L were transferred into COC1/DDP cells by lipofectin.The inhibition rate of transferred cells to cisplatin was detected by methyl thiazol tetrazolium （MTT） colori-metric assay,and compared with that in untransferred COC1/DDP cells.And the altered expression of GSTπ was also detected by RT-PCR.Results （1） GSTπ was overexpressed in cisplatin resistant cell line of COC1/DDP as compared with COC1 cell line.（2） In comparison with COC1/DDP cells,GSTπ exression in GSTπ-ASODN transfected cells was significantly decreased to 0.058 ± 0.022 and resistance index （RI） was 3.61 （P 〈 0.05）.On the contrary,missense oligodeoxynucleotide had no effect on inhibition of the expression of GSTπ （P 〉 0.05）,and the drug sensitivity of COC1/DDP cells either （P 〉 0.05）.Conclusion （1） The overexpression of GSTπ may play an important role in the induction of resistance of COC1/DDP cells to cisplatin; （2） GSTπ-ASODN can effectively reverse the resistance of COC1/DDP cells to cisplatin and increase their drug sensitivity in a sequence specific manner.

关 键 词：卵巢肿瘤 顺铂 反义寡核苷酸 谷胱甘肽转移酶π基因 耐药性 

分 类 号：R737.31[医药卫生—肿瘤]