Objective To investigate whether AZD1152 (AZD),the selective inhibitor of aurora kinase B,may play a role in the treatment of cisplatin-resistant ovarian carcinoma when administrated alone or in combination with cisplatin.Methods Hey (cisplatin-resistant ovarian cancer cell line) cells were analyzed.According to the treatment plan,Hey cells were divided into four groups (AZD group,cisplatin group,AZD + cisplatin group and control group).Methyl thiazolyl tetrazolium (MTT) assay was used to test the cells proliferation,caspase-3/7 activity analysis was used to analyze cells apoptosis,and fluorescence insitu hybridization (FISH) assay was used to determine the copy the number of chromosome 7 and checked the copy numbers of hTERC gene and C-myc gene.Results MTT test showed that proliferation of AZD group was lower than that in control group(P ＜ 0.01).The cells proliferation with the treatment with 10 and 20 nmol/L AZD for 24 hours was (81.4 ± 3.6)％ and (81.4 ± 3.6)％ respectively,and the cells proliferation for 48 hours was (43.1 ± 2.0) ％ and (38.5 ± 1.6) ％ respectively,which was significantly lower than control group (100％,P ＜ 0.01) ; Treated with the same concentration of AZD,inhibition of proliferation was significantly enhanced as the time extended (P ＜ 0.01).Proliferation in group AZD+cisplatin was lower than that in cisplatin group (P ＜ 0.01) which suggest that there were additive effects after combined AZD with cisplatin.Compared with control group,caspase-3/7 activity in AZD group increased significantly (P =0.000),and the same results was seen between AZD ± cisplatin group and cisplatin group or AZD group (all P ＜ 0.01).Compared with cisplatin group or control group,the copy numbers of hTERC,C-myc and the number of chromosome were significantly increased in AZD group and AZD + cisplat group (all P ＜ 0.05).Conclusions AZD could inhibite ovarian cancer cells proliferation and induce cells apoptosis significantly.AZD alone or in combination with cisplatin may result in the increased cells polyploidy.%目的 探讨极光激酶B选择性抑制剂AZD1152(AZD)对顺铂耐药卵巢上皮性癌(卵巢癌)细胞的作用.方法 以顺铂耐药卵巢癌细胞株Hey细胞为研究对象,实验分为4组,AZD组、顺铂组、AZD+顺铂组、对照组,四甲基偶氮唑蓝(MTT)比色法检测4组Hey细胞的相对增殖率[以吸光度(A)药物组/A对照组×100％表示],生物荧光法检测4组Hey细胞的凋亡水平[以半胱氨酸天冬氨酸蛋白酶(caspase) 3/7相对活性表示],荧光原位杂交(FISH)技术检测4组Hey细胞中hTERC和C-myc癌基因的表达[hTERC(红色)、C-myc(绿色)的荧光点数分别表示hTERC和C-myc基因的拷贝数]及Hey细胞的染色体倍数[SE7(蓝色)的荧光点数表示染色体的倍体数].结果 MTT比色法检测显示,AZD组(10和20 nmol/L的AZD分别处理24和48 h)Hey细胞的相对增殖率[10、20 nmoL/L的AZD处理24 h时分别为(81.4±3.6)％、(81.4±3.6)％,处理48 h时分别为(43.1±2.0)％、(38.5±1.6)％],均明显低于对照组的100％ (P ＜0.01)；且同一浓度的AZD处理时,随着处理时间(分别为24、48 h)的延长,其增殖抑制作用明显增强(P＜0.01).与顺铂组(2、4、7μmol/L的顺铂分别处理24和48 h)相比,AZD+顺铂组(10或20 nmoL/L的AZD +2、4或7μmol/L的顺铂分别处理24和48 h)Hey细胞相对增殖率明显下降(P＜0.01),而且AZD和顺铂两药间存在叠加效应.生物荧光法检测显示,AZD组(10 nmol/L) Hey细胞中caspase-3/7的相对活性为1.45 ±0.08,与对照组(1.00)比较,差异有统计学意义(P=0.000)；AZD+顺铂组(10 nmol/L的AZD+4μmol/L的顺铂)、顺铂组(4 μmol/L) Hey细胞中caspase-3/7的相对活性分别为1.76 ±0.12、1.01 ±0.04,AZD+顺铂组分别与顺铂组、AZD组比较,差异均有统计学意义(P＜0.01).FISH技术检测显示,与对照组、顺铂组相比,AZD组和AZD+顺铂组细胞中hTERC、C-myc的拷贝数和Hey细胞的染色体倍数均明显增加,差异均有统计学意义(P＜0.05).结论 AZD可以有效抑制顺铂耐药卵巢癌Hey细胞的增殖,并诱导细胞凋亡.AZD单用或与顺铂联合使用均可诱导Hey细胞多倍体的形成.
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