CDCA3在肾癌中的表达及其对肾癌细胞增殖与凋亡的影响

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

目的:探讨细胞周期相关因子3(CDCA3)在肾癌组织中的表达水平及其对肾癌细胞增殖和凋亡的影响。方法:回顾性分析来宾市人民医院2012年1月1日—2015年1月1日收治86例肾癌患者临床病理资料,采用实时荧光定量聚合酶链反应(qRT-PCR)方法分析CDCA3在肾癌组织和癌旁组织中表达水平;分析CDCA3在肾癌组织中表达水平与预后关系;肾癌细胞系(786-O)中转染si-CDCA3及阴性对照;MTT法检测细胞增殖能力变化;流式细胞仪检测细胞周期和凋亡变化;Westernblot检测CDCA3、CyclinD1和cleaved caspase-3蛋白的表达变化。结果:CDCA3在肾癌组织中表达水平高于对应癌旁组织[(1.66±0.13) vs.(1.07±0.16),t=4.137,P<0.05]且表达水平与预后相关(P<0.05);肾癌细胞中转染si-CDCA3和si-NC,CDCA3 Mrna[(0.97±0.33) vs.(1.27±0.40),t=2.137,P<0.05]和蛋白表达下调[(0.45±0.12) vs.(0.22±0.16),t=2.455,P<0.05];沉默CDCA3,G;/G;比例增加[(61.34±0.42) vs.(50.57±0.38),t=9.367,P<0.05]、肾癌细胞凋亡率增加[(21.42±2.36) vs.(48.64±3.53),t=5.016,P<0.05]、CyclinD1蛋白表达下调[(0.62±0.25) vs.(0.44±0.09),t=2.349,P<0.05]、cleaved caspase-3蛋白表达上调[(0.35±0.10) vs.(0.46±0.50),t=2.943,P<0.05]。结论:CDCA3 mRNA在肾癌组织中高表达,可能通过靶向CyclinD1和cleaved caspase-3蛋白调控肿瘤细胞增殖和凋亡。

Objective: To investigate the expression level of cell division cycle associated protein 3(CDCA3) in renal carcinoma tissues and its effect on proliferation and migration of renal carcinoma cells. Methods: The clinicopathological data of 86 renal cancer patients admitted to Laibin People’s Hospital from January 1 st, 2012 to January 1 st, 2015 were retrospectively analyzed. QRT-PCR was used to analyze the relative expression levels of CDCA3 mRNA in cancer tissues and adjacent tissues. The relationship between the expression level of CDCA3 in renal carcinoma tissues and the prognosis of patients was also analyzed. Renal cell lines(786-O) were transfected with SI-CDCA3 and negative control. MTT assay was used to detect the change of cell proliferation ability. Cell cycle and apoptosis were detected by flow cytometry. Western blot detected the expression changes of CDCA3, CyclinD1 and cleaved caspase-3 proteins. Results: CDCA3 expression level in renal carcinoma tissues was higher than that in adjacent tissues [(1.66±0.13) vs.(1.07±0.16), t=4.137, P<0.05], and the expression level was correlated with prognosis(P<0.05). Si-CDCA3 mRNA [(0.97±0.33) vs.(1.27±0.40), t=2.137, P<0.05] and protein expression was down-regulated [(0.45±0.12) vs.(0.22±0.16), t=2.455, P<0.05] after transfection with Si-CDCA3 and Si-NC in renal carcinoma cells. CDCA3 silence, the G;/G;ratio increased [(61.34±0.42) vs.(50.57±0.38), t=9.367, P<0.05], the apoptosis rate of renal carcinoma cells increased [(21.42±2.36) vs.(48.64±3.53), t=5.016, P<0.05], and the expression of CyclinD1 protein was down-regulated [(0.62±0.25) vs.(0.44±0.09), t=2.349, P<0.05] and cleaved caspase-3 expression was up-regulated [(0.35±0.10) vs.(0.46±0.50), t=2.943, P<0.05]. Conclusion: CDCA3 mRNA is highly expressed in renal carcinoma tissues, which may regulate tumor cell proliferation and apoptosis by targeting CyclinD1 and cleaved caspase-3 proteins.