FAT1通过调节CDK4／CDK6／CCND1复合物表达抑制食管鳞癌细胞增殖  

FAT1 inhibits cell proliferation of esophageal squamous cell carcinoma through regulating the expression of CDK4/CDK6/CCND1 complex

作　　者：胡晓玲[1] 翟元芳 李国栋[4] 邢建峰[3] 杨洁 毕炀辉 王娟 师如意 

机构地区：[1]山西医科大学基础医学院药理教研室,太原030001 [2]山西医科大学转化医学研究中心,太原030001 [3]山西医科大学机能实验室,太原030001 [4]山西省人民医院耳鼻喉头颈外科,太原030012

出　　处：《中华肿瘤杂志》2018年第1期14-20,共7页Chinese Journal of Oncology

基　　金：国家自然科学基金（81602176）;山西省青年科学基金（201601D202107、201701D221249）;山西医科大学博士启动基金（03201006）;山西医科大学青年基金（057487）;山西医科大学331基础医学科技培植基金（201219）

摘　　要：目的探讨食管鳞癌患者中FATl的表达情况及其对细胞生长的调控机制。方法收集食管鳞癌组织标本制作组织芯片，采用免疫组化检测FATl在食管鳞癌及癌旁正常组织中的表达。在高表达FATl食管鳞癌细胞系YSE2和Col0680N中，应用慢病毒感染法干扰FATI后，采用四甲基偶氮唑蓝法和平板克隆形成实验观察FATl对细胞生长的影响。流式细胞术检测敲低FATl的表达对细胞周期和凋亡率的影响。Westernblot检测敲低FATI的表达对细胞周期标志物表达的影响。结果FATl在食管鳞癌及癌旁正常组织中的相对表达水平分别为66．97±21．53和78．13±16．76，差异有统计学意义（P〈0．05）。敲低FATl表达的细胞生长速度加快，克隆形成能力增强。在YSE2细胞中，对照组细胞分裂1次的时间为（1570±51）min，敲低FATl表达后细胞分裂时间为（1356±31）min；在Col0680N细胞中，对照组细胞分裂1次时间为（1532±53）min，敲低FATl表达后细胞分裂时间为（1290±30）min，差异均有统计学意义（P〈0．05）。敲低FATl表达促进细胞周期由G，期向S期转化，上调cDK4／cDK6／ccNDl复合物的表达。结论FATl通过调节食管鳞癌细胞CDK4／CDK6／CCNDl复合物的表达，抑制细胞周期由G．期向s期转化，抑制细胞增殖。Objective To explore the expression of FAT1 in esophageal squamous cell carcinoma （ESCC） tissues, and its effect on cell proliferation. Methods The expression levels of FAT1 protein in human ESCC tissues and matched adjacent normal tissues were determined by immunohistochemistry （IHC）. Lentivirus based knockdown of FAT1 was carried out in YSE2 and Colo6gON cell lines and 3-（ 4,5-dimethyl- 2-thiazolyl）-2,5-diphenyl-2H tetrazolium bromide （MTT） assays was performed to examine the effect of FAT1 on the proliferation of these ESCC cells. Colony formation assay was used to detect the colony formation ability. Flow eytomet17 was performed to analyze the cell cycle and apoptosis. The expression levels of cell cycle markers in FAT1 knock out ESCC cell lines were detected by real-time quantitative reverse transcription polymerase chain reaction （qRT-PCR） and Western blot. Results The relative expression of FAT1 in ESCC tissues was 66.97±21.53, significantly lower than 78.13± 16.76 of adjacent normal tissues （P〈0.05）. Knockdown of FATI promoted cell proliferation and colony formation. In YSE2 cell, the division time in negative control （NC） group was （1 570±51 ） rain, significantly longer than （1 356±31 ） rain inshFAT1 group. In Colo680N cell, division time in NC group was （ 1 532±53） min, significantly longer than （1 290±30） min in shFAT1 group （P〈0.05）. Knockdown of FAT1 promoted Gl-to S-phase transition and resulted in the upregulation of CDK4/CDK6/CCND1. Conclusion FAT1 inhibits the proliferation and G1- to S-phase transition of ESCC cells through regulating the protein expression of CDK4/CDK6/CCND1 complex.

关 键 词：食管肿瘤 癌 鳞状细胞 FAT1 细胞增殖 细胞周期 

分 类 号：R735.1[医药卫生—肿瘤]