塞来昔布对乳腺癌MCF-7细胞的增殖抑制作用及对BCRA1、Caspase-3、p53表达的影响

Inhibitory effect of celecoxib on proliferation of breast cancer MCF-7 cells and its influence on BCRA1,Caspase-3,p53 expressions in vitro

目的探讨塞来昔布对乳腺癌MCF-7细胞的增殖抑制作用及对BCRA1、Caspase-3、p53表达的影响。方法乳腺癌MCF-7细胞培养后,取对数生长期细胞用于实验。根据加入塞来昔布终浓度的不同分为实验四组:实验A组(20μmol/L)、实验B组(40μmol/L)、实验C组(80μmol/L)和实验D组(160μmol/L);以加入0.1%DMSO为正常对照组。采用MTT法检测塞来昔布对MCF-7细胞的体外抑制作用;采用DAPI染色法测定细胞凋亡形态;采用流式细胞仪检测塞来昔布对MCF-7细胞凋亡率的影响;采用Western blot(WB)法检测BCRA1、Caspase-3、p53的表达。结果 MTT法检测结果:实验A、B、C、D组抑制率明显高于正常对照组,且随塞来昔布浓度增加抑制率递升(P<0.01);半数抑制浓度(IC50)=91.3628μmol/L。DAPI染色法测定结果:正常对照组细胞完整,未发现细胞凋亡;实验B、C、D组经处理后出现细胞生长抑制及细胞体积缩小现象,DAPI染色后见明显的细胞核固缩和断裂,且细胞凋亡现象随塞来昔布浓度增加而明显。流式细胞仪检测结果:实验B、C、D组凋亡率明显高于对照组,且随塞来昔布浓度增加而递增(P<0.01);WB法检测结果:实验B、C、D组BCRA1、Caspase-3、p53蛋白的相对表达量明显高于正常对照组,且随塞来昔布浓度的增加BCRA1、Caspase-3、p53蛋白的相对表达量依次上调(P<0.01)。结论塞来昔布可明显抑制乳腺癌MCF-7细胞的增殖,其作用机制可能与上调BCRA1、Caspase-3、p53的表达相关。

Objective To investigate the inhibitory effect of celecoxib on proliferation of breast cancer cells（ MCF-7 cell）and its influence on BCRA1,Caspase-3,p53 expressions. Methods After being cultured,MCF-7 cells in logarithmic growth phase were used for experiment. The MCF-7 cells were divided into four experiment groups according to the different final concentration of celecoxib（ which being added） ： experiment group A（ 20 μmol / L）,experiment group B（ 40 μmol /L）,experiment group C（ 80 μmol / L）,experiment group D（ 160 μmol / L）,and the MCF-7 cells added 0. 1% DMSO were served as control group. MTT method was used to detect the inhibitory effect of celecoxib on proliferation of MCF-7 cells in vitro. DAPI staining method was used to observe the morphology of cell apoptosis. Flow cytometry was used to detect the influence of celecoxib on MCF-7 cells apoptosis rate. Western blot method was used to detect BCRA1,Caspase-3 and p53 proteins expressions. Results The result of MTT method detection showed that the inhibition rates in experiment groups A,B,C and D were all significantly higher than that in control group,and progressively increased with the increase of celecoxib concentrations（ P〈0. 01）,and the 50% inhibitory concentration（ IC50） was 91. 3628 μmol/L. The result of DAPI staining showed that（ 1） in control group,the cells were complete,and cell apoptosis was not found.（ 2） in experiment groups B,C and D,the phenomena of cell growth inhibition and cell volume reduction appeared after treatment; obvious nuclear condensation and fracture can be seen after DAPI staining; the cell apoptosis phenomenon increased obviously with the increase of celecoxib concentrations. The result of flow cytometry detection showed that the cell apoptosis rates in experi-ment groups B,C and D were significantly higher than that in control group,and progressively increased with the increase of celecoxib concentrations（ P〈0. 01）. The result of Western blot method detection showed that in experiment groups B,C and D,the relative expression quantities of BCRA1,Caspase-3 and p53 proteins were significantly higher than those in control group,and were up-regulated in turn with the increase of celecoxib concentrations（ P〈0. 01）. Conclusion Celecoxib can obviously inhibit the proliferation of MCF-7 cells,and the mechanism may be related to the up-regulation of BCRA1,Caspase-3 and p53 expressions.