戊地昔布联合阿霉素对人乳腺癌细胞增殖的抑制作用及其机制研究

Inhibitory Effects of Valdecoxib Combined with Adriamycin on Proliferation of Human MCF-7 Cells and Its Mechanism

目的:研究戊地昔布联合阿霉素对人乳腺癌细胞(MCF-7)增殖的抑制作用及其机制。方法:将人MCF-7细胞随机分为空白对照组、溶剂对照组、阿霉素(0.25mg·L-1)组、戊地昔布(25、50、100μmol·L-1)组及联用组(阿霉素0.25mg·L-1+戊地昔布25、50、100μmol·L-1),加入相应药物继续培养,检测培养24、48、72h(n=6)后各组人MCF-7细胞的增殖抑制率;检测戊地昔布25μmol·L-1时培养48h后各组细胞周期和增殖细胞核抗原(PCNA)蛋白、细胞周期蛋白(CyclinD1)及环氧酶2(COX-2)蛋白表达。结果:与溶剂对照组比较,除戊地昔布25μmol·L-1培养24h外,阿霉素组、戊地昔布组及联用组增殖抑制率均明显增加(P均<0.01),且呈浓度、时间依赖性;与阿霉素组和戊地昔布组比较,联用组增殖抑制率均明显增加(P均<0.01)。戊地昔布组细胞阻滞于G0/G1期,S期细胞比例明显减少(P<0.05);联用组细胞阻滞于G0/G1期和G2/M期。与溶剂对照组比较,戊地昔布组、阿霉素组及联用组PCNA、CyclinD1蛋白表达均明显降低(P<0.05或P<0.01),仅戊地昔布组和联用组COX-2蛋白表达明显降低(P<0.05或P<0.01)。结论:戊地昔布与阿霉素联用具有协同抑制人MCF-7细胞增殖的作用,可能与抑制CyclinD1和PCNA蛋白表达从而阻滞细胞周期有关。

OBJECTIVE： To study the inhibition effects of valdecoxib combined with adriamycin on the proliferation of human MCF-7 ceils and its mechanism. METHODS： Human MCF-7 cells were randomly divided into blank control group, solvent control group, adriamycin group （0.25 mg.L-1）, valdecoxib group （25, 50, 100 μmol·L^-1） and combination group （adriamycin 0.25 mg. L^-1＋valdecoxib 25, 50, 100 μmol·L^-1）. The inhibition rates of human MCF-7 cell proliferation were determined 24, 48, 72 h af-ter treated with relevant medicine （n=6）. Cell cycle, the protein expressions of PCNA, Cyclin D1 and COX-2 in various groups were detected 48 h after cultured in valdecoxib 25 μmol·L^-1. RESULTS： Compared with sdvent control group, the inhibition rate of cell proliferation was increased significantly in adriamycin group, valdecoxib group and combination group, except for culture of valdecoxib 25 μmol·L^-1 for 24 h （all P〈0.01）, in concentration and time-dependant manner; compared with adriamycin group and valdecoxib group, the inhibition rate of cell proliferation was increased significantly in combination group （all P〈0.01）. Valde-coxib markedly blocked the MCF-7 cell progression by arresting the ceils in the G0/G1 phase, and the percentage of cells in S phase decreased dramatically （P〈0.05）; and valdecoxib combined with adriamycin blocked the MCF-7 cell progression by arresting the cells in GdGI and GJM phase （P〈0.05）. Compared with solvent control group, the protein expressions of PCNA and Cyclin D1 in valdecoxib group, adriamycin group and combination group were decreased significantly （P〈0.05 or P〈0.01）, while the protein expression of COX-2 in valdecoxib group and combination group decreased significantly （P〈0.05 or P〈0.01）. CONCLUSION： Valdecoxib synchronized with adriamycin can inhibit the proliferation of human MCF-7 cells significantly, which may be associated with the inhibition of Cyclin D1 and PCNA expression and the blockage of cell cycle.