CAJ | 학술논문

目的观察X连锁凋亡抑制蛋白(XIAP)小分子抑制剂Embelin在体外安全剂量下能否逆转耐药.方法 CCK-8法检测Embelin对亲本株和耐药株增殖的影响及单用培美曲塞和联合Embelin分别对两株细胞的半数抑制浓度(IC50)变化.逆转录-聚合酶链反应(RT-RCR)、Western blot 检测Embelin作用亲本株和耐药株前后XIAP mRNA和蛋白的表达.流式细胞仪分析在耐药细胞中单用培美曲塞组和联合Embelin组凋亡差异.结果 Embelin均能抑制耐药株和亲本株增殖,具浓度依赖性,在浓度＜2.5 mg/L时对细胞基本无不良反应.单用培美曲塞和联合Embelin只在耐药株IC50差异有统计学意义(P＜0.05).Embelin作用亲本株和耐药株后耐药株XIAP mRNA和蛋白分别下凋33%、25%(P＜0.05).Embelin流式细胞分析在耐药细胞中联合Embelin 组凋亡比例高于单用培美曲塞组(25.87±2.81)%、(38.73±2.55)%,(P＜0.05).结论 XIAP在耐药株中表达增高,Embelin能降低其在耐药株中的表达并提高胰腺癌Patu8988耐药细胞株对培美曲塞的敏感性,对亲本珠影响不大.
목적 관찰X련쇄조망억제단백(XIAP)소분자억제제Embelin재체외안전제량하능부역전내약.방법 CCK-8법검측Embelin대친본주화내약주증식적영향급단용배미곡새화연합Embelin분별대량주세포적반수억제농도(IC50)변화.역전록-취합매련반응(RT-RCR)、Western blot 검측Embelin작용친본주화내약주전후XIAP mRNA화단백적표체.류식세포의분석재내약세포중단용배미곡새조화연합Embelin조조망차이.결과 Embelin균능억제내약주화친본주증식,구농도의뢰성,재농도＜2.5 mg/L시대세포기본무불량반응.단용배미곡새화연합Embelin지재내약주IC50차이유통계학의의(P＜0.05).Embelin작용친본주화내약주후내약주XIAP mRNA화단백분별하조33%、25%(P＜0.05).Embelin류식세포분석재내약세포중연합Embelin 조조망비례고우단용배미곡새조(25.87±2.81)%、(38.73±2.55)%,(P＜0.05).결론 XIAP재내약주중표체증고,Embelin능강저기재내약주중적표체병제고이선암Patu8988내약세포주대배미곡새적민감성,대친본주영향불대.
Objective To investigate whether safe doses of Embelin, a small-molecular weight inhibitor of X-linked inhibitor of apoptosis protein (XIAP) can reverse the resistance of human pancreatic carcinoma Patu8988 cell line to pemetrexed. Methods The effect of Embelin on growth inhibition of pemetrexed-resistant Patu8988 strain and its parent strain and half maximal inhibitory concentration ( IC50 ) of pemetrexed alone and its combination with Embelin for the growth of the two cell strains were determined by the cell counting Kit-8 assay. Reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting were used to detect the changes in the expression of XIAP mRNA and protein after the interference of Embelin. Results Embelin presented growth inhibition on both cell stains without significant differences in vitro in a dose-dependent manner. The maximal safe dose of Embelin for both strains was 2. 5 mg/L. Only in Patu8988 strain to pemetrexed was noted a significant difference in the IC50 value between pemetrexed alone and pemetrexed in combination with safe doses of Embelin. The expression of XIAP mRNA and protein was decreased by 33% and 25% respectively (P＜0. 05) in pemetrexed-resistant Patu8988 strain after the interference of Embelin. Flow cytometry analysis showed that the apoptosis rate of drug-resistant cells treated with pemetrexed in combination with Embelin was higher than that of cells treated with pemetrexed alone [ (38.73±2.55)% vs (25. 87 ±2.81)%, P＜0.05]. Conclusion Embelin can reduce the the higher expression of XIAP mRNA and protein levels in drug-resistant cells and increase the sensitivity of drug-resistant Patu8988 cell strain to pemetrexed, but has no significant effect on parental Patu8988 strain.