细胞周期相关因子3在乳腺癌组织中的表达及对乳腺癌细胞增殖的影响

Expression of CDCA3 in breast cancer tissues and its effect on the proliferation of breast cancer cells

目的 检测乳腺癌组织中细胞周期相关因子3(cell division cycle associated protein 3,CDCA3)的表达水平,分析CDCA3对乳腺癌细胞生长增殖的影响。方法 对我院乳腺癌标本库中41例不同分期乳腺癌组织进行免疫组化检测,结合生物信息学方法与癌症基因组图谱(TCGA)数据库,分析CDCA3在乳腺癌组织中的表达情况。利用shRNA构建稳定敲低CDCA3的乳腺癌MCF7细胞株,观察下调CDCA3后乳腺癌细胞的增殖情况。采用蛋白免疫印迹法检测增殖相关NF-кB信号转导通路相关蛋白NF-кB1(p50)、RelA(p65)表达水平,观察CDCA3对NF-кB通路的调控作用。结果 免疫组化及生物信息学分析结果均显示乳腺癌组织中CDCA3蛋白表达量(1.83±0.72)显著高于癌旁组织(1.02±0.51),且CDCA3高表达患者的总体生存率(59.24%)显著低于CDAC3低表达者(70.15%),差异均有统计学意义(P<0.05)。CDCA3表达下调后,MCF7细胞增殖受抑制,G1期细胞比例显著增加,NF-кB1(p50)、RelA(p65)表达水平降低,差异均有统计学意义(P<0.05)。GSEA结果表明,CDCA3可能通过调节G1S检查点、mTOR信号通路、E2F信号通路、NF-кB信号通路、胆固醇平衡、糖酵解反应、细胞的有丝分裂纺锤体等对乳腺癌的发生发展产生影响。结论 CDCA3在乳腺癌肿瘤组织中高表达,且其表达水平与乳腺癌预后呈负相关,干扰CDCA3表达可抑制乳腺癌细胞的增殖。

Objective To detect the expression level of cell division cycle associated protein 3(CDCA3) in breast cancer tissues and analyze the effect of CDCA3 on the growth and proliferation of breast cancer cells.Methods Immunohistochemical detection was performed on 41 breast cancer tissues of different stages in the breast cancer specimen bank of our hospital,and the expression of CDCA3 in breast cancer tissues was analyzed by combining bioinformatics methods with the cancer genome map(TCGA) database.A breast cancer MCF7 cell line stably knockdown CDCA3 was constructed using shRNA,and the proliferation of breast cancer cells after down-regulation of CDCA3 was observed.The expression levels of proliferation related NF-кB signal transduction pathway-related proteins NF-кB1(p50) and RelA(p65) were detected by Western blotting,and the regulatory effect of CDCA3 on the NF-кB pathway was explored.Results Immunohistochemical and bioinformatics analysis showed that the expression of CDCA3 protein in breast cancer tissues(1.83±0.72) was significantly higher than that in adjacent tissues(1.02±0.51),and the overall survival rate of patients with high expression of CDCA3(59.24%) was significantly lower than that of patients with low expression of CDAC3(70.15%).The differences were statistically significant(P<0.05).After the down-regulation of CDCA3 expression,the proliferation of MCF7 cells was inhibited,the proportion of cells in G1 phase was significantly increased,and the expression levels of NF-кB1(p50) and RelA(p65) were decreased.The differences were statistically significant(P<0.05).GSEA results indicated that CDCA3 might affect the occurrence and development of breast cancer by regulating G1 S checkpoints,mTOR signal pathway,E2 F signal pathway,NF-кB signal pathway,cholesterol balance,glycolysis reaction,cell mitotic spindle,etc.Conclusions CDCA3 is highly expressed in breast cancer tissues,and its expression level is negatively correlated with the prognosis of breast cancer.Interfering with the expression of CDCA3 can inhibit the proliferation of breast cancer cells.