葡萄糖、棕榈酸通过ROCK1-KIF2A通路诱导结肠癌细胞中心体扩增  

Glucose and Palmitic Acid Induce Colon Cancer Cell Centrosome Amplification through ROCK1-KIF2A Pathway

作　　者：王朴 杜斌 牛紫旭 周飞伦 李少钦 WANG Pu;DU Bin;NIU Zixu;ZHOU Feilun;LI Shaoqin(Department of Cell Biology,Faculty of Basic Medicine,Changzhi Medical College,Changzhi 046000,China;Department of Parasite,Faculty of Basic Medicine,Changzhi Medical College,Changzhi 046000,China;Second Clinical College,Changzhi Medical College,Changzhi 046000,China;School of Life Science,Jiangsu Normal University,Xuzhou 211006,China)

机构地区：[1]长治医学院,基础医学部,生物教研室,长治046000 [2]长治医学院,基础医学部,寄生虫教研室,长治046000 [3]长治医学院,第二临床学院,长治046000 [4]江苏师范大学,生命科学学院,徐州211006

出　　处：《中国细胞生物学学报》2021年第8期1581-1587,共7页Chinese Journal of Cell Biology

基　　金：山西省科技创新项目(批准号:2020L0373)资助的课题。

摘　　要：为了研究微管解聚酶KIF2A如何参与葡萄糖、棕榈酸诱导的结肠癌HCT116细胞中心体扩增,该实验使用葡萄糖(Glu,25 mmol/L)、棕榈酸(Pal,150μmol/L)处理HCT116细胞后,再使用ROCK1抗体免疫沉淀其相互作用蛋白,并将沉淀结果通过高通量质谱HPLC/MS鉴定。通过Western blot、siRNA、免疫荧光验证ROCK1的相互作用蛋白。结果显示,在葡萄糖、棕榈酸共同处理后,中心体定位的ROCK1可结合中心体蛋白定位的微管解聚酶KIF2A。敲降KIF2A的表达可降低葡萄糖、棕榈酸诱导的中心体扩增率。生物信息学分析显示,癌组织KIF2A的mRNA表达水平较健康组织显著增加(健康组织3.275±0.385,StageⅠ4.460±0.682,StageⅡ4.436±0.620,StageⅢ4.365±0.680,StageⅣ4.442±0.555,P<0.01),不同临床分期的结肠癌组织中KIF2A的表达量无显著差异。KIF2A的低表达与结肠癌的不良预后呈正相关(中位生存期为低表达1765天vs高表达3042天)。综上所述,ROCK1-KIF2A通路在葡萄糖、棕榈酸诱导的中心体扩增中起到了关键作用,为糖尿病相关的结肠癌综合防治提供了实验依据。This work was to investigate how microtubule depolymerase KIF2A was involved in centrosome amplification induced by glucose and palmitic acid in colon cancer HCT116 cells.HCT116 cells were co-treated with glucose (25 mmol/L) and palmitic acid (150 μmol/L),and then immunoprecipitated with ROCK1 antibody,and the precipitation results were identified by high throughput mass spectrometry HPLC/MS.Western blot,siRNA and immunofluorescence were used to verify the interacting proteins of ROCK1.The results showed that the centrosomal located ROCK1 could bind to the centrosomal protein-located microtubule depolymerase KIF2A after treatment with glucose and palmitic acid.Knockdown of KIF2A expression could reduce the amplification rate of centrosomes induced by glucose and palmitic acid.Bioinformatics analysis showed that the mRNA expression level of KIF2A in cancer tissue was significantly higher than that in healthy tissue (healthy tissues 3.275±0.385,Stage I 4.460±0.6818,Stage II 4.436±0.620,Stage III 4.365±0.680,Stage IV 4.442±0.555,P<0.01),there is no difference in the expression of KIF2A in colon cancer tissues of different clinical stages.The low KIF2A expression was positively correlated with poor prognosis of colon cancer (median survival:low expression 1 765 days vs high expression 3 042 days,P<0.01).In conclusion,ROCK1-KIF2A pathway plays a key role in centrosome amplification induced by glucose and palmitic acid,which provides experimental basis for comprehensive prevention and treatment of diabetes-related colon cancer.

关 键 词：葡萄糖 棕榈酸 中心体扩增 ROCK1 KIF2A 

分 类 号：R735.35[医药卫生—肿瘤]