PARP-1抑制剂PJ-34对多药耐药骨髓瘤细胞株的影响以及与FA／BRCA途径的关系  

Effect of PARP-1 inhibitor P J-34 on multi-drug resistance in human multiple myeioma cell line and its relationship with FA/BRCA pathway

作　　者：熊婷[1] 魏恒[1] 陈小琼[1] 肖晖[1] 

机构地区：[1]武汉大学中南医院血液内科,430071

出　　处：《中华医学遗传学杂志》2014年第3期312-316,共5页Chinese Journal of Medical Genetics

基　　金：国家自然科学基金（81001053）;中央高校专项科研基金武汉大学自主项目（4101041）

摘　　要：目的研究PARP-1抑制剂PJ-34对人多发性骨髓瘤多药耐药细胞株RPM18226／R的作用，探讨PJ-34对DNA损伤修复和耐药性的影响以及与FA／BRCA途径的联系。方法应用CCK-8比色法检测细胞抑制率；应用实时荧光定量PCR检测参与FA／BRCA途径对DNA损伤修复的基因表达的变化；采用Western免疫印迹法检测FA／BRCA途径中的关键蛋白FANCD2的表达；用Annexin V-FITC／PI双标法流式细胞术检测PJ-34对细胞凋亡的影响；用彗星实验检测PJ-34对细胞DNA损伤修复的影响。结果PJ-34能够显著增加RPMl8226／R细胞对马法兰的敏感性，使马法兰的IC50值由20．43t2mol／L降至7．8μmol／L；PJ-34能够抑制DNA损伤的修复，且与抑制FA／BRCA途径有关；PJ-34协同马法兰能促进对RPMI8226／R细胞的致凋亡作用。结论PJ-34能够显著增强RPMI8226／R细胞对马法兰的敏感性，通过抑制FA／BRCA途径对DNA损伤的修复来逆转细胞的耐药性，因此，PJ-34可能在克服多发性骨髓瘤多药耐药的治疗中具有一定的临床价值。Objective To investigate the effect of PARP-1 inhibitor PJ-34 on multi-drug resistance in a human multiple myeloma cell line and its connection with FA/BRCA pathway in DNA damage repair. Methods A CCK-8 assay was used to measure the inhibition rate. Real-time quantitative PCR was used to detect expression changes of DNA repair genes involved in the FA/BRCA pathway. Western blotting assay was used to detect expression of key protein FANCD2 in the FA/BRCA pathway. Annexin V-FITC/PI double staining flow cytometry was used to measure cell apoptosis induced by PJ-34. A COMET assay was used to detect the effect of PJ-34 on DNA damage repair. Results PJ-34 could significantly enhance the sensitivity of RPMI8226/R cells to melphalan. The IC50 value of melphalan was dropped from 20.43μmol/ L to 7.8μmol/L. PJ-34 could inhibit the DNA damage repair, and the effect was related with the inhibition of FA/BRCA pathway. PJ-34 and melphalan showed a synergistic effect in promoting the apoptosis of RPMI8226/R cells. Conclusion PJ-34 can reverse the resistance of RPMI8226/R cells to melphalan by inhibiting the FA/BRCA pathway, which in turn can induce suppression of DNA damage repair. Therefore, PJ-34 may have clinical value in overcoming the multi-drug resistance of multiple myeloma.

关 键 词：多发性骨髓瘤 多药耐药 DNA损伤 FA BRCA途径 

分 类 号：R733.3[医药卫生—肿瘤]