阿司匹林和塞来昔布对乳腺癌细胞MCF-7中COX-2及VEGF表达的影响

目的：通过研究非甾体类抗炎药物（NSAIDs）阿司匹林和塞来昔布对乳腺癌细胞MCF-7增殖及环氧化酶-2（COX-2）和血管内皮生长因子（VEGF）表达的影响，探讨NSAIDs的抗肿瘤作用。方法：MCF-7细胞根据所加药物不同分为不同浓度的阿司匹林（2.5、5.0和10.0mmol·L^-1）组和塞来昔布（30、60和120 μmol·L^-1）组，以不含药物的培养液作为阴性对照组。MTT法检测不同时间（24、48和72h）各组细胞增殖抑制率，应用免疫组织化学SP法检测不同时间（24和48h）各组MCF-7细胞中COX-2及VEGF的表达。结果：①2.5 mmol·L^-1阿司匹林作用48h、30 μmol·L^-1塞来昔布作用48和72 h及60 μmol·L^-1塞来昔布作用48 h细胞增殖抑制率与阴性对照组比较差异无显著性（P>0.05），其他各剂量组细胞增殖抑制率高于阴性对照组（P<0.05）。10.0 mmol·L^-1阿司匹林作用24、48和72 h细胞增殖抑制率高于同一时间2.5 mmol·L^-1阿司匹林组（P<0.05）。120 μmol·L^-1塞来昔布作用24、48和72 h细胞增殖抑制率高于同一时间30和60 μmol?L^-1塞来昔布组（P<0.05）。2.5、5.0和10.0 mmol?L^-1阿司匹林作用72 h较作用24 h细胞增殖抑制率升高（P<0.05）。②MCF-7细胞中均有COX-2和VEGF蛋白表达，均定位于细胞浆，染色呈黄或棕黄色。③30 μmol·L^-1塞来昔布作用24、48 h和60 μmol·L^-1塞来昔布作 48 h与阴性对照组比较MCF-7中COX-2和VEGF表达差异无显著性（P>0.05），其他各剂量组细胞中COX-2和VEGF表达均低于阴性对照组(P<0.05)。10.0 mmol?L^-1阿司匹林作用24和48 h细胞中COX-2和VEGF表达低于同一时间2.5和5.0 mmol·L^-1阿司匹林组（P<0.05）。120 μmol·L^-1塞来昔布作用24和48 h细胞中COX-2和VEGF表达低于同一时间30 μmol·L^-1塞来昔布组（P<0.05）。10.0 mmol·L^-1阿司匹林和120 μmol·L^-1塞来昔布作用48 h较作用24 h细胞中COX-2和VEGF表达降低（P<0.05）。结论：阿司匹林和塞来昔布均有抑制乳腺癌细胞MCF-7增殖的作用,同时亦能抑制细胞中COX-2和VEGF的表达，上述抑制作用随药物浓度的增加、作用时间的延长而增强。

Effects of aspirin and celecoxib on expressions of COX-2 and VEGF in breast cancer cells MCF-7

Objective To study the anti-tumor effect of aspirin and celecoxib on breast cancer cells MCF-7 through investigating their effects on the expressions of COX-2,VEGF and proliferation of MCF-7 cells.Methods The human breast cancer cells MCF-7 were divided into 2.5,5.0 and 10.0 mmol·L-1 aspirin groups and 30,60 and 120 μmol·L-1 celecoxib groups,the MCF-7 cells without treatment were used as negative control group.The inhibitory rates were detected by MTT assay after MCF-7 cells were treated for 24,48,72 h.The expressions of COX-2 and VEGF in MCF-7 cells after treated for 24 and 48h were detected by immunohistochemical assay.Results ① There were no significant differences of the inhibitory rates of the MCF-7 cell line between 2.5 mmol·L-1 aspirin group(48 h),30 μmol·L-1 celecoxib group(48 and 72 h),60 μmol·L-1 celecoxib group(48 h) and control group(P>0.05),the inhibitory rates in the other groups were increased compared with control group(P<0.05).The inhibitory rates of MCF-7 cells in 10.0 mmol·L-1 aspirin group after treated for 24,48 and 72 h were higher than those of 2.5 mmol·L-1 aspirin group at the same time(P<0.05).The inhibitory rates of MCF-7 cells in 120 μmol·L-1 celecoxib group after treated for 24,48 and 72 h were higher than those in 30 μmol·L-1 or 60 μmol·L-1 celecoxib groups at the same time(P<0.05).The inhibitory rates of MCF-7 cells in 2.5,5.0 and 10.0 mmol·L-1 aspirin groups after treated for 72 h were higher than those of MCF-7 cells after treated for 24 h(P<0.05).②The expressions of COX-2 and VEGF located in MCF-7 cytoplasm with yellow and pale brown staining.③There were no significant differences of the expressions of COX-2 and VEGF in MCF-7 cells between 30 μmol·L-1 celecoxib group(24,48 h),60 μmol·L-1 celecoxib group(48 h) and negative control group(P>0.05),the expressions of COX-2 and VEGF in MCF-7 cells in the other groups were lower than that in negative control group(P<0.05).The expressions of COX-2 and VEGF in MCF-7 cells treated with 10.0 mmol·L-1 aspirin for 24 and 48 h were decreased compared with those in 2.5 or 5.0 mmol·L-1 aspirin groups at the same time(P<0.05).The expressions of COX-2 and VEGF in MCF-7 cells treated with 120 μmol·L-1 celecoxib for 24 and 48 h were decreased compared with those in 30 μmol·L-1 celecoxib group at the same time(P<0.05).The expressions of COX-2 and VEGF in MCF-7 cells treated with 10.0 mmol·L-1 aspirin or 120 μmol·L-1 celecoxib for 48 h were decreased compared with those of MCF7 cells treated for 24 h(P<0.05).Conclusion Aspirin and celecoxib have inhibitory effects on the growth of human breast cancer cells MCF-7,and they can inhibit the expressions of COX-2 and VEGF in MCF-7 cells in a dose and time dependent manner.