塞来昔布联合多西他赛抑制人乳腺癌MDA-MB-231细胞增殖和诱导凋亡的作用研究  

Celecoxib and Docetaxel Inhibited Proliferation and Induced Apoptosis of Breast Cancer Cell Line MDA-MB-231

作　　者：许燕艳[1] 翟凤钰[2] 王亚帝[3] 岳莉[4] 漆起贵[1] 王芳[1] 

机构地区：[1]辽宁医学院,辽宁省锦州市121001 [2]濮阳市油田总医院放疗科 [3]辽宁医学院附属第三医院肿瘤科 [4]辽宁医学院附属第一医院肿瘤科,辽宁省锦州市121001

出　　处：《中国全科医学》2013年第9期1004-1007,1011,共5页Chinese General Practice

基　　金：CSCO-金港榄香烯肿瘤学研究生课题基金(z-2011-004)

摘　　要：目的探讨塞来昔布联合多西他赛抑制人乳腺癌MDA-MB-231细胞增殖和诱导凋亡的作用。方法人乳腺癌MDA-MB-231细胞分别给予塞来昔布60μmol/L(塞来昔布组)、多西他赛2.0μmol/L(多西他赛组)、60μmol/L塞来昔布+2.0μmol/L多西他赛(联合用药组)处理,培养24、48、72 h。四甲基偶氮唑蓝微量酶反应比色法(MTT法)检测细胞存活率;瑞氏-吉姆萨染色观察细胞形态学变化;Annexin-V/PI双染流式细胞术分析细胞凋亡情况;蛋白印迹法(Western Blot)检测bcl-2、bax、survivin的表达。结果塞来昔布组、多西他赛组和联合用药组不同时间MDA-MB-231细胞细胞存活率比较,差异有统计学意义(F组间=9.78,P=0.03,F时间=10.45,P=0.03)。联合用药组作用于细胞24 h、48 h、72 h后相互作用系数(CDI)分别为:(0.46±0.21)、(0.23±0.14)、(0.17±0.04)。瑞氏-吉姆萨染色400倍光镜下观察到塞来昔布组和多西他赛组细胞出现双核及停滞于分裂中期,联合用药组细胞表现为染色质固缩、胞核碎裂及凋亡小体等典型的凋亡形态。药物作用于细胞48 h后,塞来昔布组细胞凋亡率为(20.14±1.22)%,多西他赛组为(54.60±1.37)%,联合用药组为(59.85±1.42)%,差异有统计学意义(F=1 395.96,P=0.00)。Western Blot结果显示,塞来昔布组、多西他赛组和联合用药组作用于细胞48 h后,bcl-2表达分别下降至对照组的89.02%、75.63%、45.35%,差异有统计学意义(χ2=13.77,P<0.05);bax表达上调至对照组的136.35%、157.24%、173.89%,差异有统计学意义(χ2=4.79,P<0.05);survivin表达下降至对照组的80.42%、54.28%、35.85%,差异有统计学意义(χ2=10.28,P<0.05)。结论塞来昔布可协同多西他赛诱导乳腺癌MDA-MB-231细胞凋亡,其机制可能与下调bcl-2、survivin及上调bax蛋白有关。Objective To investigate the effect of celecoxib and docetaxel on the proliferation and apoptosis of breast cancer cell line MDA - MB - 231. Methods Breast cancer cell line MDA - MB - 231 was treated by 60μmol/L celecoxib （ cele- coxib group）, 2.0 μmol/L docetaxel （ docetaxel group） and 60μmoL/L celecoxib plus 2. 0 μmol/L docetaxel （ combination group） respectively. The cell line was cultivated for 24 h, 48 h and 72 h respectively. MTT method was used to detect cell survival rate, Wright - Giemsa staining was used to observe morphological changes, Annexin - V/PI flow cytometry was used to analyze cell apoptosis and Western Blot method was used to detect the expression of bcl - 2, bax and survivin. Results The cell survival rate of MDA - MB -231 cell line between celecoxib group, docetaxel group and combination group showed statistically significant difference （Fbetween==9.78, P=0.03, Ftime =10.45, P=0.03） .The CDI was （0.46±0.21）, （0.23±0.14） and （0. 17±0. 04） respectively after 24 h, 48 h and 72 h action on cells in the combination group. In Wright - Giemsa staining, under 400 times lens, ceils in celecoxib group and docetaxel group showed double nucleuses and stopped in metaphase, while ceils in combination group showed chromatin condensation, fragmented nucleus and apoptosis body. After 48 h action on cells, the apoptosis rate of celecoxib group, docetaxel group and combination group was （20. 14±1.22） %, （ 54. 60±1.37 ） % and （59. 85±1.42） %, and the difference was statistically significant （ F = 1 395.96, P = 0. 00） . The results of Western Blot showed that after 48 h action on cells, the expression of bcl - 2 in celecoxib group, docetaxel group and combination group de- creased to 89. 02% , 75.63% and 45.35% of the control group respectively, and the difference was statistically significant （X2 = 13.77, P 〈 0.05 ） . The expression of bax increased to 136. 35%, 157.24% and 173.89% of the control group and the difference was statistically significant

关 键 词：塞来昔布 多西他赛 乳腺肿瘤 细胞凋亡 

分 类 号：R979.1[医药卫生—药品] R737.9[医药卫生—药学]