CAJ | 학술논문

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.
목적 관찰순박내약란소암세포주COC1/DDP중곡광감태전이매π(GSTπ)기인적표체,탐토기반의과핵감산대COC1/DDP순박내약적역전작용.방법 설계합성침대GSTπ적류대탈양과핵감산(ASODN),이200 nmol/L농도전염COC1/DDP세포,용새서람법관찰순박대세포억제솔적변화,반전록-취합매련반응(RT-PCR)검측기인재mRNA수평표체적개변.결과 (1)COC1/DDP세포중GSTπ기인정고표체,기mRNA상대표체치위0.78±0.13.이COC1세포중무표체.(2)여COC1/DDP세포조비교,GSTπ-ASODN조GSTπ-mRNA강저,상대표체치위0.058±0.022,내약지수(RI)강저위3.61,차이균유통계학의의(P＜0.05);착의과핵감산조,GSTπ표체급RI개변차이무통계학의의(P＞0.05).결론 (1)GSTπ기인고표체여COC1세포획득순박내약성유밀절관계.(2) GST-ASODN능유효역전COC1/DDP세포주대순박적내약성.
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.