不同浓度硼替佐米对K562/DNR细胞株NF-κB,IκB及P-gp表达的影响

Effects of different concentrations of bortezomib on the expression of NF-κB,IκB and P-gp of DNR-resistant K562 cells

目的:观察不同浓度硼替佐米(商品名万珂,PS-341)对柔红霉素(DNR)诱导的K562耐药细胞株(K562/DNR)核因子-κB(NF-κB)、抑制蛋白κB(IκB)及P-糖蛋白(P-gp)表达的影响,探讨PS-341逆转耐药的分子机制.方法:四甲基偶氮唑盐微量酶反应比色法(MTT法)进行耐药细胞株和PS-341细胞毒性的判定.以100mg/LDNR单用或联合应用0.4,4,40μg/LPS-341作用于K562/DNR36h,检测各组NF-κb,IκB及P-gp表达情况,并测定NF-κB活性,检测各组细胞凋亡率.结果:与阴性对照组相比,DNR可诱导NF-κB表达上调、IκB表达下调、P-gp表达上调;加用PS-341可显著抑制NF-κB表达,同时使IκB表达增加、P-gp表达下降,上述作用随PS-341浓度增加而增强.NF-κB活性(%)检测示:DNR组为25.9±2.5,DNR+0.4μg/LPS-341组为20.3±2.0,DNR+4μg/LPS-341组为6.1±2.5,DNR+40μg/LPS-341组为4.6±1.6,加入PS-341后,NF-κB活性受抑,该作用随PS-341浓度增加而增强.细胞凋亡率(%)检测结果示:DNR组为22.5±4.6,DNR+0.4μg/LPS-341组为31.0±5.2,DNR+4μg/LPS-341组为43.6±7.7,DNR+40μg/LPS-341组为56.0±9.3,加入PS-341后可提高DNR引起的细胞凋亡率,该作用随PS-341浓度的增加而增强.结论:PS-341可逆转白血病细胞耐药,该作用呈浓度依赖性.

AIM ： To observe the effects of bortezomib（ PS-341 ） on the expression of NF-κB, IκB and P-gp of K562 cells induced by dannorubicin （K562/DNR）. METHODS ： MTT method was used to determine the drug-resistant K562 cells and the cellular toxicity of bortezomib; Detect the expression of NF-κB, IκB and P-gp of K562/DNR 36 h after receiving the treatment of 100 mg/L DNR only or added with 0.4,4 and 40 μg/L bortezomib. Detect the apoptosis rate in each group respectively and the activity of NF-κB was detected by ELISA method. RESULTS： Compared with the control group, the expressions of NF-κB and P-gp in K562/DNR group could be induced by DNR. When K562/DNR cells were cultured with bortezomib,the expressions of NF-κB and P-gp induced by DNR were significantly suppressed and IκB was increased, these effects increased with the increase of the concentration of PS-341. The results of NF-κBg activity showed that the team of DNR was 25. 9 ± 2. 5, DNR ± 0.4 μg/L PS-341 was 20.3 ±2.0,DNR ±4 μg/L PS-341was 6.1 ±2.5,DNR ±40 μg/L PS-341 was 4.6 ± 1.6. The activity of NF-κB was suppressed when PS-341 was added in. The results of apoptosis rate （ % ） showed that the team of DNR was 22.5 ± 4.6, DNR ± 0.4 μg/L PS-341 was 31.0 ±5.2,DNR ±4 μg/L PS-341was 43.6 ±7.7, DNR ± 40 μg/L PS-341 was 56.0 ± 9.3. The apoptosis rate increased when K562/DNR cells were cultured with bortezomib, these effects increased with the increase of concentration of bortezomib. CONCLUSION： Bortezomib could convert the cellular drug resistance and this effect showed the characteristic of concentration-dependent pattern.