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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 the proliferation of breast cancer MCF-7 cells and its effect on the expression of BCRA1, Caspase-3 and p53. Methods Breast cancer MCF-7 cells were cultured and logarithmic growth phase cells were used for the experiment. According to the difference of the final concentrations of celecoxib, the rats were divided into four groups: experimental group A (20μmol / L), experimental group B (40μmol / L), experimental group C 80μmol / L and experimental group D 160μmol / ); Adding 0.1% DMSO as the normal control group. The inhibitory effect of celecoxib on MCF-7 cells was detected by MTT assay. The morphological changes of apoptotic cells were detected by DAPI staining. The effect of celecoxib on the apoptosis rate of MCF-7 cells was detected by flow cytometry. Western blot (WB) was used to detect the expression of BCRA1, Caspase-3 and p53. Results The results of MTT assay showed that the inhibition rates in groups A, B, C and D were significantly higher than those in normal control group (P <0.01), and the IC50 values were 91.3628μmol / L. The results of DAPI staining showed that the cells in the normal control group were intact and apoptosis was not found. After the treatment, the cell growth inhibition and the cell volume shrinkage were observed in the groups B, C and D, obvious nuclear pyknosis and fragmentation were observed after DAPI staining, And the phenomenon of apoptosis with celecoxib concentration increased significantly. The results of flow cytometry showed that the apoptotic rate of group B, C and D were significantly higher than that of control group (P <0.01), and the concentration of celecoxib increased (P <0.01) The relative expression of BCRA1, Caspase-3 and p53 protein in group B was significantly higher than that in normal control group. The relative expression of BCRA1, Caspase-3 and p53 protein increased with the increase of celecoxib concentration (P <0.01). Conclusion Celecoxib can significantly inhibit the proliferation of breast cancer MCF-7 cells, which may be related to the up-regulation of the expression of BCRA1, Caspase-3 and p53.