中华妇产科杂志
中華婦產科雜誌
중화부산과잡지
CHINESE JOUNAL OF OBSTETRICS AND GYNECOLOGY
2013年
1期
46-50
,共5页
卵巢肿瘤%细胞系,肿瘤%磷酸酯类%喹唑啉类%顺铂%抗药性,肿瘤
卵巢腫瘤%細胞繫,腫瘤%燐痠酯類%喹唑啉類%順鉑%抗藥性,腫瘤
란소종류%세포계,종류%린산지류%규서람류%순박%항약성,종류
Ovarian neoplasms%Cell line,tumor%Phosphoric acid esters%Quinazolines%Cisplatin%Drug resistance,neoplasm
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.
目的 探討極光激酶B選擇性抑製劑AZD1152(AZD)對順鉑耐藥卵巢上皮性癌(卵巢癌)細胞的作用.方法 以順鉑耐藥卵巢癌細胞株Hey細胞為研究對象,實驗分為4組,AZD組、順鉑組、AZD+順鉑組、對照組,四甲基偶氮唑藍(MTT)比色法檢測4組Hey細胞的相對增殖率[以吸光度(A)藥物組/A對照組×100％錶示],生物熒光法檢測4組Hey細胞的凋亡水平[以半胱氨痠天鼕氨痠蛋白酶(caspase) 3/7相對活性錶示],熒光原位雜交(FISH)技術檢測4組Hey細胞中hTERC和C-myc癌基因的錶達[hTERC(紅色)、C-myc(綠色)的熒光點數分彆錶示hTERC和C-myc基因的拷貝數]及Hey細胞的染色體倍數[SE7(藍色)的熒光點數錶示染色體的倍體數].結果 MTT比色法檢測顯示,AZD組(10和20 nmol/L的AZD分彆處理24和48 h)Hey細胞的相對增殖率[10、20 nmoL/L的AZD處理24 h時分彆為(81.4±3.6)％、(81.4±3.6)％,處理48 h時分彆為(43.1±2.0)％、(38.5±1.6)％],均明顯低于對照組的100％ (P ＜0.01)；且同一濃度的AZD處理時,隨著處理時間(分彆為24、48 h)的延長,其增殖抑製作用明顯增彊(P＜0.01).與順鉑組(2、4、7μmol/L的順鉑分彆處理24和48 h)相比,AZD+順鉑組(10或20 nmoL/L的AZD +2、4或7μmol/L的順鉑分彆處理24和48 h)Hey細胞相對增殖率明顯下降(P＜0.01),而且AZD和順鉑兩藥間存在疊加效應.生物熒光法檢測顯示,AZD組(10 nmol/L) Hey細胞中caspase-3/7的相對活性為1.45 ±0.08,與對照組(1.00)比較,差異有統計學意義(P=0.000)；AZD+順鉑組(10 nmol/L的AZD+4μmol/L的順鉑)、順鉑組(4 μmol/L) Hey細胞中caspase-3/7的相對活性分彆為1.76 ±0.12、1.01 ±0.04,AZD+順鉑組分彆與順鉑組、AZD組比較,差異均有統計學意義(P＜0.01).FISH技術檢測顯示,與對照組、順鉑組相比,AZD組和AZD+順鉑組細胞中hTERC、C-myc的拷貝數和Hey細胞的染色體倍數均明顯增加,差異均有統計學意義(P＜0.05).結論 AZD可以有效抑製順鉑耐藥卵巢癌Hey細胞的增殖,併誘導細胞凋亡.AZD單用或與順鉑聯閤使用均可誘導Hey細胞多倍體的形成.
목적 탐토겁광격매B선택성억제제AZD1152(AZD)대순박내약란소상피성암(란소암)세포적작용.방법 이순박내약란소암세포주Hey세포위연구대상,실험분위4조,AZD조、순박조、AZD+순박조、대조조,사갑기우담서람(MTT)비색법검측4조Hey세포적상대증식솔[이흡광도(A)약물조/A대조조×100％표시],생물형광법검측4조Hey세포적조망수평[이반광안산천동안산단백매(caspase) 3/7상대활성표시],형광원위잡교(FISH)기술검측4조Hey세포중hTERC화C-myc암기인적표체[hTERC(홍색)、C-myc(록색)적형광점수분별표시hTERC화C-myc기인적고패수]급Hey세포적염색체배수[SE7(람색)적형광점수표시염색체적배체수].결과 MTT비색법검측현시,AZD조(10화20 nmol/L적AZD분별처리24화48 h)Hey세포적상대증식솔[10、20 nmoL/L적AZD처리24 h시분별위(81.4±3.6)％、(81.4±3.6)％,처리48 h시분별위(43.1±2.0)％、(38.5±1.6)％],균명현저우대조조적100％ (P ＜0.01)；차동일농도적AZD처리시,수착처리시간(분별위24、48 h)적연장,기증식억제작용명현증강(P＜0.01).여순박조(2、4、7μmol/L적순박분별처리24화48 h)상비,AZD+순박조(10혹20 nmoL/L적AZD +2、4혹7μmol/L적순박분별처리24화48 h)Hey세포상대증식솔명현하강(P＜0.01),이차AZD화순박량약간존재첩가효응.생물형광법검측현시,AZD조(10 nmol/L) Hey세포중caspase-3/7적상대활성위1.45 ±0.08,여대조조(1.00)비교,차이유통계학의의(P=0.000)；AZD+순박조(10 nmol/L적AZD+4μmol/L적순박)、순박조(4 μmol/L) Hey세포중caspase-3/7적상대활성분별위1.76 ±0.12、1.01 ±0.04,AZD+순박조분별여순박조、AZD조비교,차이균유통계학의의(P＜0.01).FISH기술검측현시,여대조조、순박조상비,AZD조화AZD+순박조세포중hTERC、C-myc적고패수화Hey세포적염색체배수균명현증가,차이균유통계학의의(P＜0.05).결론 AZD가이유효억제순박내약란소암Hey세포적증식,병유도세포조망.AZD단용혹여순박연합사용균가유도Hey세포다배체적형성.
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.
