PBK/TOPK通过改变糖酵解关键酶表达水平增加结直肠癌细胞放射敏感性  

PBK/TOPK increases the radiosensitivity of colorectal cancer cells by changing the expression levels of key glycolytic enzymes

作　　者：郭佳星 王振堂 林宇[1] 王雨哲 额尔德木图[1] 李静[1] 郁志龙[1] 宝莹娜[1] GUO Jiaxing;WANG Zhentang;LIN Yu;WANG Yuzhe;Eerdemutu;LI Jing;YU Zhilong;BAO Yingna(Department of Radiation Therapy,Affiliated Hospital of Inner Mongolia Medical University,Inner Mongolia Hohhot 010050,China)

机构地区：[1]内蒙古医科大学附属医院放疗科,内蒙古呼和浩特010050

出　　处：《现代肿瘤医学》2022年第7期1167-1171,共5页Journal of Modern Oncology

基　　金：内蒙古自治区自然科学基金(编号:2019MS08124);内蒙古医科大学青年创新基金(编号:YKD2017QNCX074)。

摘　　要：目的:探讨在结直肠癌细胞中PDZ结合激酶(PDZ binding kinase,PBK)/T-淋巴因子激活的杀伤细胞来源的蛋白激酶(T-cell-originated protein kinase,TOPK)能否通过影响糖酵解关键酶表达改变放射敏感性。方法:通过慢病毒转染敲低LS174T细胞中的PBK/TOPK表达水平。利用克隆形成实验检测细胞的放射敏感性,Real time-PCR和Western blot检测PBK/TOPK、葡萄糖转运蛋白1(glucose transporter 1,GLUT1)和己糖激酶(hexokinase 2,HK2)在RNA水平和蛋白水平的表达差异。结果:与对照组比较,敲低PBK/TOPK的表达后细胞克隆形成集落较小较少。通过Western blot和Real time-PCR实验发现:单纯给予8 Gy照射后,细胞中GLUT1相对表达量至48 h最低,72 h呈现升高趋势;HK2相对表达量则呈现完全相反的变化,在48 h表达量最高;PBK/TOPK单纯敲低组,GLUT1相对表达量至48 h最低,72 h呈现升高趋势,HK2相对表达量则呈现完全相反的变化,48 h表达量最高;敲低PBK/TOPK联合8 Gy照射组中,GLUT1相对表达量在48 h内呈现升高趋势,并于48 h达到顶峰,72 h出现降低趋势,而HK2相对表达量的变化为随着时间逐渐降低。结论:敲低PBK/TOPK能够增强结直肠癌细胞放射敏感性,其机制可能与糖酵解关键酶GLUT1及HK2的表达呈现完全相反的变化有关。Objective:To explore whether PBK/TOPK has effect on the radiosensitivity by affecting the expression of key glycolytic enzymes in colorectal cancer cells.Methods:Lentivirus transfection was used to knocked down PBK/TOPK in LS174T cells.It was tested the radiosensitivity by cloning formation.The expression of PBK/TOPK,GLUT1 and HK2 were detected by Real time-PCR and Western blot.Results:The cloning formation of LS174T cells with knocking down PBK/TOPK was smaller than the control group.It showed that GLUT1 in LS174T cells with 8 Gy radiation alone decreased to the minimal expression at 48 h,but HK2 had an opposite expression that was highest expression at 48 h.Besides it was found that the expression of GLUT1 decreased in PBK/TOPK knocking down group and then showed an upward trend,the expression of HK2 showed a completely opposite change,with the highest expression at 48 h.GLUT1 showed an increasing trend within 48 h and reached a peak at 48h,then a decreasing trend appeared in the cell with PBK/TOPK knocking down combined with 8 Gy radiation,while the expression level of HK2 gradually decreased with time.Conclusion:Knockdown of PBK/TOPK can enhance the radiosensitivity of colorectal cancer cells.The mechanism may be related to the completely opposite changes in the expression of key glycolytic enzymes GLUT1 and HK2.

关 键 词：PBK/TOPK 糖酵解 葡萄糖转运蛋白1 己糖激酶2 放射敏感性 

分 类 号：R735.3[医药卫生—肿瘤]