CDCA3在食管癌中的表达及其对食管癌细胞增殖与凋亡的影响

Expression of CDCA3 in esophageal carcinoma and its effect on proliferation and apoptosis of esophageal cancer cells

目的研究细胞周期相关因子(CDCA3)在食管癌组织和细胞中的表达及其对食管癌细胞增殖与凋亡的影响。方法实时荧光定量PCR(qPCR)检测50例食管癌组织及其配对的癌旁组织中CDCA3 mRNA的表达水平,同时检测CDCA3 mRNA在不同食管癌细胞系(Eca109、Kyse-170、TE1)和人食管鳞状上皮细胞系Het-1A中的表达水平;采用si-CDCA3-1、si-CDCA3-2转染Eca109和TE1细胞,另转染si-NC作为阴性对照,MTS法检测细胞增殖能力,流式细胞仪检测细胞周期和凋亡变化;Western blot检测CDCA3、CyclinD1和cleaved caspase-3蛋白的表达变化。结果食管癌组织中CDCA3 mRNA的表达高于癌旁组织(P<0.05),Eca109、Kyse-170和TE1细胞中CDCA3 mRNA表达高于人食管鳞状上皮细胞(P<0.05)。沉默CDCA3表达后,Eca109细胞和TE1细胞的增殖率降低,G0/G1期细胞比例增加(P<0.05)。沉默CDCA3表达后,Eca109细胞和TE1细胞的凋亡率增加(P<0.05)。沉默CDCA3表达可降低CyclinD1蛋白表达和增加cleaved caspase-3蛋白表达(P<0.05)。结论CDCA3 mRNA在食管癌组织和细胞系中均高表达,干扰CDCA3的表达可能通过降低CyclinD1蛋白表达抑制细胞增殖,增加cleaved caspase-3蛋白表达促进细胞凋亡,提示CDCA3可能为食管癌的潜在治疗靶标。

Objective To study the expression of cell cycle related factor(CDCA3)in esophageal carcinoma tissues and cells and its effect on proliferation and apoptosis of esophageal cancer cells.Methods Real-time quantitative PCR(qPCR)was used to detect the expression of CDCA3 mRNA in 50 cases of esophageal cancer tissues and their adjacent paracancerous tissues,and to detect CDCA3 mRNA in different esophageal cancer cell lines(Eca109,Kyse-170,TE1)and human esophageal squamous epithelial cell het-1A.Eca109 and TE1 cells were transfected with si-CDCA3-1 and si-CDCA3-2,and si-NC was used as a negative control.Cell proliferation was detected by MTS assay,and cell cycle and apoptosis were detected by flow cytometry.Western blot assay was used to detect the expression changes of CDCA3,CyclinD1 and cleaved caspase-3 proteins.Results The expression of CDCA3 mRNA in esophageal carcinoma tissues was higher than that in adjacent tissues(P<0.05).The expression of CDCA3 mRNA in Eca109,Kyse-170 and TE1 cells was higher than that in normal human esophageal squamous cells(P<0.05).After silencing CDCA3 expression,the proliferation rate of Eca109 cells and TE1 cells decreased,and the proportion of cells in G0/G1 phase increased(P<0.05).After silencing CDCA3 expression,the apoptosis rate of Eca109 cells and TE1 cells increased(P<0.05).Silencing CDCA3 expression reduced CyclinD1 protein expression and increased cleaved caspase-3 protein expression(P<0.05).Conclusion CDCA3 mRNA is highly expressed in esophageal carcinoma tissues and cell lines.Interfering with the expression of CDCA3 may inhibit cell proliferation by decreasing the expression of CyclinD1 protein,and increase the expression of cleaved caspase-3 protein to promote apoptosis.CDCA3 may be a potential therapeutic target for esophageal cancer.