驱动蛋白家族蛋白14在食管鳞癌组织的表达及其对食管鳞癌细胞增殖和周期的影响

Expression of kinesin family protein 14 in esophageal squamous cell carcinoma and its effects on cell proliferation and cell cycle

目的 观察驱动蛋白家族蛋白14（KIF14）在食管鳞癌组织的表达,探讨其表达下调对食管鳞癌细胞增殖和周期的影响.方法 采用免疫组织化学检测87例食管鳞癌组织和对应的正常食管上皮组织中KIF14的表达,分析其表达与临床病理学参数的关系.利用Western blot检测不同食管鳞癌细胞及正常食管上皮细胞中KIF14蛋白的表达,将KIF14小干扰RNA（siRNA）和对照siRNA转染食管鳞癌Eca109细胞,利用Western blot检测转染后KIF14蛋白表达的变化.进一步采用细胞计数试剂盒（CCK-8）和流式细胞术检测KIF14表达下调对食管鳞癌Eca109细胞增殖和周期的影响.结果 KIF14在食管鳞癌组织中表达的阳性率（62.3%）显著高于正常食管上皮组织（21.8%）,差异有统计学意义（χ2=30.474,P=0.000）.KIF14蛋白的表达与食管鳞癌患者的组织学分级、临床分期以及淋巴结转移均密切相关（P=0.000、0.026、0.007）,但与食管鳞癌患者的性别和年龄均无明显相关（P=0.821、0.644）.此外,KIF14在食管鳞癌细胞中的表达水平显著高于正常食管上皮细胞Het-1A,并且KIF14 siRNA能显著下调食管鳞癌Eca109细胞中KIF14蛋白的表达.最为重要的是KIF14表达下调显著抑制食管鳞癌Eca109细胞的增殖,静止细胞周期在G0/G1期.结论 KIF14在食管鳞癌的发生发展中发挥重要作用.

Objective To investigate the expression of Kinesin family protein 14 （KIF14） in esophageal squamous cell carcinoma （ESCC）,and explore its effects of its downregulation on cell proliferation and cell cycle.Methods Immunohistochemistry was used to investigate the expression of KIF14 in 87 cases of ESCC tissues and paired normal esophageal tissues,and the correlations between KIF14 expression and clinic pathological features were analyzed.Western blotting was employed to detect the expression KIF14 protein in various ESCC cells and normal esophageal cells,and KIF14 small interfering RNA （siRNA） and control siRNA were utilized to transfect to ESCC Eca109 cells,and expression of KIF14 protein was examined after transfection.Furthermore,cell counting kit-8 （CCK-8） and Flow cytometry were used to investigate the effects of KIF14 downregulation on cell proliferation and cell cycle of ESCC Eca109 cells.Results The positive expression ratio of KIF14 in ESCC tissues （62.3%） was significantly higher than that in normal esophageal tissues （21.8%）,and the differences were statistical significance （χ2=30.474,P=0.000）.KIF14 protein expression was tightly associated with histological differentiation,clinic staging and lymph node metastasis （P=0.000,0.026,0.007）,but not related to gender and age （P=0.821,0.644）.In addition,expression of KIF14 in ECSS cells was significantly higher than that in normal esophageal cell Het-1A,and KIF14 siRNA markedly downregulated the expression of KIF14 protein in Eca109 cells.Most notably,KIF14 downregulation obviously suppressed cell proliferation and arrested cell cycle in G0/G1 phase in Eca109 cells.Conclusion KIF14 may play a pivotal role in the occurrence and development of ESCC,and thus may be an underlying molecular target.