CDK14对人食管癌细胞增殖的影响及其可能机制研究  

Effect of CDK14 on Proliferation of Human Esophageal Carcinoma Cells and its Possible Mechanism

作　　者：钱佶[1] 王燏婵[2] 张学俭[1] 

机构地区：[1]江苏大学附属宜兴医院消化内科,214200 [2]南通大学免疫学与微生物学教研室

出　　处：《胃肠病学》2016年第10期585-591,共7页Chinese Journal of Gastroenterology

摘　　要：背景:CDK14是一种新发现的细胞周期蛋白依赖性激酶,在多种恶性肿瘤中表达增高,并与肿瘤恶性行为相关。目的:探讨CDK14对人食管癌细胞增殖的影响及其可能机制。方法:采用蛋白质印迹法和免疫组化法检测8例新鲜食管鳞癌组织、96例食管鳞癌石蜡包埋组织和人食管癌细胞株Eca-109中的CDK14和细胞增殖标记物PCNA、Ki-67表达,分析CDK14表达与食管癌临床病理特征和患者预后的关系。以血清饥饿释放试验分析CDK14表达与Eca-109细胞细胞周期的关系。以shRNA干扰Eca-109细胞中的CDK14表达,观察抑癌蛋白Rb磷酸化水平、细胞周期进程和细胞增殖能力的变化。结果:CDK14在食管癌组织和细胞中呈高表达,与PCNA、Ki-67的表达趋势相一致,其表达与食管癌的大小、组织学分级、浸润和转移显著相关(P<0.05),高表达者总体生存率显著低于低表达者(P<0.05)。血清饥饿释放试验显示CDK14的表达具有细胞周期依赖性。干扰CDK14可抑制Eca-109细胞Rb蛋白磷酸化,细胞发生G1期阻滞,克隆形成数显著减少(P均<0.05)。结论:CDK14在食管癌中呈高表达,其可能通过促进下游Rb蛋白磷酸化推动细胞周期进程,从而促进肿瘤细胞增殖,参与食管癌的发生、发展。Background： CDK14 is a novel cyclin-dependent kinase,which is overexpressed in a variety of cancer and related to their malignant behavior. Aims： To investigate the effect of CDK14 on proliferation of human esophageal carcinoma cells and its possible mechanism. Methods： Expressions of CDK14 and two cell proliferation markers,PCNA and Ki-67 were estimated in 8 fresh-frozen specimens of esophageal squamous cell carcinoma（ ESCC）,96 paraffin-embedded specimens of ESCC,and human ESCC cell line Eca-109 by Western blotting and immunohistochemistry. Correlations of CDK14 expression with the clinicopathological characteristics and prognosis of ESCC were analyzed. Serum starvation and release assay was performed to evaluate the relationship between CDK14 expression and cell cycle progression in Eca-109 cells.Furthermore,Eca-109 cells were transiently transfected with shRNA-CDK14 to reduce CDK14 protein level,and then the phosphorylation of tumor suppressor protein Rb,cell cycle progression and proliferation capability of Eca-109 cells were determined. Results： CDK14 was highly expressed in both ESCC tissue and cell line,which was paralleled with the expressions of PCNA and Ki-67 and correlated significantly with the tumor size,histological grade,invasiveness and metastasis of ESCC（ p〈0. 05）. The overall survival was poor in patients with high CDK14 expression than those with low CDK14 expression（ p〈0. 05）. Serum starvation and release assay showed that the expression of CDK14 was cell cycledependent. Knockdown of CDK14 reduced the expression level of phosphorylated Rb,induced significant G1 phase arrest and resulted in less colony formation in Eca-109 cells（ P all 0. 05）. Conclusions： CDK14 is highly expressed in ESCC.It may promote cell cycle progression by phosphorylating downstream Rb protein,thus enhancing the proliferation of tumor cells,and ultimately participating in the occurrence and development of ESCC.

关 键 词：细胞周期蛋白依赖性激酶14 食管肿瘤 成视网膜细胞瘤蛋白 细胞周期 细胞增殖 

分 类 号：R735.1[医药卫生—肿瘤]