酪氨酸激酶抑制剂逆转黏附诱导的骨髓瘤细胞的耐药性  

Tyrosine Kinase Inhibitor Reverses Adriamycin Resistance Mediated by Cell Adhesion in RPMI8226 Cells

作　　者：潘耀柱[1] 陈协群[1] 高广勋[1] 顾宏涛[1] 张永清[1] 董宝侠[1] 白庆咸[1] 朱华峰[1] 

机构地区：[1]第四军医大学西京医院血液内科,西安710032

出　　处：《中国实验血液学杂志》2006年第2期267-270,共4页Journal of Experimental Hematology

摘　　要：为了探讨酪氨酸激酶抑制剂STI571对RPMI8226细胞黏附、黏附诱导耐药以及靶细胞Rac1mRNA表达的影响,本研究采用结晶紫染色法分析了RPMI8226细胞与纤连蛋白(fibronectin,FN)的黏附率,用MTT法检测瘤细胞的增殖性,并用半定量RT-PCR研究Rac1mRNA水平的变化。结果显示:RPMI8226细胞与FN黏附1、6、12小时,其黏附率分别为(43.71±2.18)%、(55.63±1.56)%、(63.42±2.46)%;用20μmol/LSTI571处理1、6、12小时后黏附率分别降为(15.12±1.04)%、(17.58±1.32)%、(17.24±1.59)%;组间差异显著(P<0.05);阿霉素作用后,FN黏附组细胞IC50值[(1.46±0.04)μmol/L]显著高于BSA组[(0.78±0.03)μmol/L](P<0.05);FN联合STI571组IC50值[(0.81±0.05)μmol/L]与BSA联合STI571组[(0.74±0.02)μmol/L]相比无显著性差异(P>0.05),但却低于FN组(P<0.05);半定量RT-PCR显示,Rac1mRNA水平在20μmol/LSTI571处理14小时后明显下降。结论:STI571能降低RPMI8226细胞与FN的黏附率、逆转黏附介导的阿霉素耐药,并且可以下调其Rac1mRNA水平。To study the effects of tyrosine-kinase inhibitor STI571 on the adhesion of RPMI8226 cells to fibronectin （FN）, cell adhesion mediated adriamycin-resistance and the Racl mRNA expression, the adhesion of RPMI8226 cells to fibronectin and drug resistance mediated by cell adhesion were determined by means of crystal violet staining and MTF assays respectively, Racl mRNA levels in RPMI8226 cells were examined by semi-quantitative RT-PCR. The results showed that STI571 could inhibit the adhesion of RPMI8226 cells to fibronectin. When RPMI8226 cells had been adhered to FN or BSA-coated wells for 1,6 and 12 hours, the adhesion rates were （43.71 ±2.18）% ,（55.63 ± 1.56）%, and （63.42 ± 2.46）% respectively. After treatment with STI571 20 μmol/L, the adhesion rates decreased to （ 15.12 ± 1.04）%, （17.58 ±1.32）% and （17.24 ±1.59）% respectively （P〈0.05）. The experiment revealed that growth of RPMI8226 cells adhered to FN-coated plates had a significant advantage over growth on BSA-coated plates when exposed to adriamycin （Adr） for 1 hour followed by a 24-hour culture period, and the mean IC50 value for FN-adhered cells was （ 1.46 ± 0.04 ） μmol/L while mean IC50 value for BSA control was （ 0.78 ± 0.03 ） μmol/L （ P 〈 0.05 ）. Following treatment with 20 μmol/L STI571, the mean IC50 values for FN and BSA adhered cells were （0.81 ±0.05 ）μmol/L, （0.74 ±0.02）μmol/L respectively, there was no significant difference between them（ P 〉 0.05 ）. RT-PCR demonstra- ted that the relative Racl mRNA level （Racl/GAPDH） in RPMI8226 cells was downregulated following being treated with 20μmol/L STI571. It is concluded that STI571 can inhibit the adhesion of RPMI8226 cells to fibronectin, reverse cell adhesion mediated adriamycin-resistance, and downregulate Racl mRNA level.

关 键 词：酪氨酸激酶抑制剂 STI571 骨髓瘤细胞 RPMI8226细胞 黏附 耐药 

分 类 号：R733.3[医药卫生—肿瘤]