摘要
目的探讨比卡鲁胺(BIC)联合化疗药物紫杉醇(PTX)对雄激素受体(AR)阳性的三阴性乳腺癌MDA-MB-231细胞的增殖抑制作用及可能的作用机制。方法采用CCK-8试剂盒观察不同浓度的BIC(0.1、1.0、10.0μmol/L)和PTX(0.1、1.0、10.0、100.0、1 000.0、10 000.0 nmol/L)以单药及不同联合给药方式处理后,对MDA-MB-231细胞增殖的抑制作用。细胞增殖抑制率比较采用单因素方差分析。组间两两比较采用LSD法。选取10 nmol/L PTX及10 nmol/L DMSO分别处理MDA-MB-231细胞样品(各3个)72 h,采用生物信息学方法分析样品的相关基因表达芯片数据,采用校正t检验筛选出差异基因。结果使用不同浓度的BIC分别处理MDA-MB-231细胞24、48、72 h后,各组MDA-MB-231细胞增殖抑制率在不同时间点差异均有统计学意义(F=4.124、8.189、4.139,P=0.037、0.004、0.032)。BIC 10.0μmol/L组MDA-MB-231细胞增殖抑制率在48 h最高,为(12.9±5.5)%。不同浓度的PTX分别处理MDA-MB-231细胞24、48、72 h后,不同浓度组MDA-MB-231细胞增殖抑制率在不同时间点差异均有统计学意义(F=8.407、47.432、14.907,P均<0.001)。PTX在48 h时对MDA-MB-231细胞的半数抑制浓度(IC50)为5 380.0 nmol/L。5 000.0 nmol/L PTX单药或联合不同浓度(0.1、1.0、10.0μmol/L)的BIC同时处理MDA-MB-231细胞48 h后,5 000.0 nmol/L PTX单药处理组与3个实验组中细胞增殖抑制率分别为(53.2±2.7)%、(53.2±3.1)%、(51.7±3.4)%、(51.0±2.3)%,组间差异无统计学意义(F=0.831,P=0.492)。采用5 000.0 nmol/L PTX和10.0μmol/L BIC以不同的序贯方式联合给药处理MDA-MB-231细胞(PTX 24 h+BIC 24 h组、BIC 24 h+PTX 24 h组、PTX 48 h+BIC 24 h组、BIC 48 h+PTX 24 h组),并用5 000.0 nmol/L PTX(PTX 48 h组)和10.0μmol/L BIC(BIC 48 h组)单药处理及同时联合给药(PTX 48 h+BIC 48 h组)分别处理MDA-MB-231细胞后,各组间细胞增殖抑制率差异有统计学意义(F=241.466,P<0.001)。其中,两两比较结果显示,PTX 24 h+BIC 24 h组细胞增殖抑制率为(72.9±1.9)%,高于BIC 24 h+PTX 24 h组的(42.9±1.7)%(P<0.001),PTX 48 h组的(60.9±3.7)%(P<0.001)和PTX 48 h+BIC 48 h组的(60.3±4.1)%(P<0.001)。PTX处理组中有EGR1、FST、FOS、IL8、IL6、RPL27A及CA2 7个基因的表达量与DMSO处理组比较,差异均有统计学意义(t=18.647、10.336、10.098、9.683、9.408、9.050、8.001,P均<0.050)。结论通过先PTX再BIC的序贯联合给药方式较单药及其他联合给药方式能够更有效抑制AR阳性三阴性乳腺癌细胞MDA-MB-231的增殖,两者间可能存在协同作用。
Objective To investigate the inhibitory effect of bicalutamide（ BIC） combined with paclitaxel（ PTX） on the proliferation of androgen receptor（ AR）-positive triple negative breast cancer MDAMB-231 cells and its possible mechanism. Methods The CCK-8 kit was used to determine the effect of BIC（ 0. 1,1. 0,10. 0 μmol/L） and PTX at different concentrations（ 0. 1,1. 0,10. 0,100. 0,1000. 0,10 000. 0 nmol/L） in monotherapy or in sequential combination of both on the proliferation of MDA-MB-231 cells. Inhibition rate was compared using one-way analysis of variance. The pairwise comparison was performed using the LSD method. MDA-MB-231 cells were treated with 10 nmol/L PTX and 10 nmol/L DMSO respectively for 72 h. Three cell samples were taken in each group to analyze the relevant gene expression profiling in array using a bioinformatic method. The adjusted t test was used to screen out differential genes.Results After MDA-MB-231 cells were treated with different concentrations of BIC for 24,48 and 72 h,respectively,the inhibition rates of MDA-MB-231 cells were statistically different at different time points（ F =4. 124,8. 189,4. 139,P = 0. 037,0. 004,0. 032）. The inhibition rate of MDA-MB-231 cells reached the highest [（ 12. 9 ± 5. 5） %] at 48 h after the treatment of 10. 0 μmol/L BIC. The inhibition rates of MDA-MB-231 cells were significantly different at different time points（ F = 8. 407,47. 432,14. 907,P0. 001） after the treatment of PTX at different concentrations. The half inhibitory concentration（ IC50） of PTX in MDA-MB-231 cells at 48 h was 5 380. 0 nmol/L. After 48 h treatment of 5 000. 0 nmol/L PTX alone or combined with 0. 1,1. 0,10. 0 μmol/L BIC,the inhibition rate of MDA-MB-231 cells was（ 53. 2 ± 2. 7） %,（ 53. 2 ± 3. 1） %,（ 51. 7 ± 3. 4） %,（ 51. 0 ± 2. 3） % in PTX monotherapy group and three experimental groups,respectively,indicating no significant difference（ F = 0. 831,P = 0. 492）. MDA-MB-231 cells were treated with sequential combination of 5 000. 0 nmol/L PTX and 10. 0 μmol/L BIC（ PTX 24 h＋BIC 24 h group,BIC 24 h＋PTX 24 h group,PTX 48 h＋BIC 24 h group,BIC 48 h ＋ PTX 24 h group）,the monotherapy with 5 000. 0 nmol/L PTX（ PTX 48 h group） or 10. 0 μmo/L BIC（ BIC 48 h group） and the synchronous combined therapy of PTX and BIC（ PTX 48 h ＋ BIC 48 h group）,respectively. The result showed that there was a statistically significant difference in inhibition rate（ F = 241. 466,P〈0. 001）. The result of pairwise comparison showed that the inhibition rate in PTX 24 h ＋ BIC 24 h group was（ 72. 9 ± 1. 9） %,significantly higher than（ 42. 9 ± 1. 7） %in BIC 24 h ＋ PTX 24 h group（ P〈0. 001）,（ 60. 9 ± 3. 7） % in PTX 48 h group（ P〈0. 001） and（ 60. 3 ±4. 1） % in PTX 48 h ＋ BIC 48 h group（ P〈0. 001）. There was a significant difference in the expression of seven genes（ EGR1,FST,FOS,IL8,IL6,RPL27 A and CA2） between PTX-treated group and DMSOtreated group（ t = 18. 647,10. 336,10. 098,9. 683,9. 408,9. 050,8. 001,all P〈0. 050） Conclusions Sequential administration of PTX and BIC can inhibit the proliferation of AR-positive triple negative breast cancer MDA-MB-231 cells more effectively compared with the monotherapy and other combination methods. The two drugs may have the synergistic effect.
作者
丁钥
许焱
丁丽
朱小泉
张永强
Ding Yue, Xu Yan, Ding Li, Zhu Xiaoquan, Zhang Yongqiang.(Department of Medical Oncology, Beijing Hospital/ National Center of Gerontology, Beijing 100730, China)
出处
《中华乳腺病杂志（电子版）》
CAS
CSCD
2018年第3期135-140,共6页
Chinese Journal of Breast Disease(Electronic Edition)
关键词
乳腺肿瘤
雄激素受体拮抗剂
化学疗法
联合药物疗法
早期生长反应蛋白质1
Breast neoplasms
Androgen receptor antagonists
Chemotherapy
Combinationmodality therapy
Early growth response protein 1
