摘要
目的探讨肿瘤微环境中肿瘤相关巨噬细胞对食管癌细胞干性的影响以及阿司匹林抗肿瘤的潜在机制。方法按照不同处理情况分组，分为对照组、ASA组、KYSE－450与M2共孵育组（KYSE－450＋M2组）、KYSE－450与M2共孵育＋ASA组（KYSE－450＋M2＋ASA组）、M2与shCCL2－KYSE450共孵育组（M2＋shCCL2－KYSE450组）、M2与shCCL2－KYSE450共孵育＋ASA组（M2＋ shCCL2－KYSE450＋ASA组）、M2与shCCL2－KYSE450共孵育组（M2＋shCCL2－KYSE450组）、M2与shCCL2－KYSE450共孵育＋ASA组（M2＋shCCL2－KYSE450＋ASA组）。采用细胞成球实验检测阿司匹林对KYSE－450成球能力的影响。采用实时荧光定量PCR、Western blot法和流式细胞术检测阿司匹林处理各组KYSE－450细胞后，KYSE－450细胞中不同趋化因子、干性基因Nanog和细胞表面干性指标CD90表达的差异。结果ASA组和KYSE－450＋M2组细胞的成球数目分别为（7.00±1.23）个和（34.33±2.33）个，与对照组[（14.50±2.33）个]比较，差异均有统计学意义（均P〈0.05）。KYSE－450＋M2＋ASA组细胞的成球数目为（20.67±2.33）个，与KYSE－450＋M2组比较，差异有统计学意义（P〈0.05）。对照组和ASA组细胞中Nanog基因的表达水平分别为1.00和0.50±0.10，差异有统计学意义（P〈0.05）。KYSE－450＋M2组和M2＋KYSE－450＋ASA组细胞中Nanog基因的表达水平分别为1.74±0.13和1.43±0.05，差异有统计学意义（P〈0.05）。M2＋shCCL2－KYSE450＋ASA组和M2＋shCCL2－KYSE450组细胞中Nanog基因的表达水平分别1.22±0.11和1.17±0.08，差异无统计学意义（P＝0.69）。流式细胞术检测结果显示，对照组和ASA组细胞中CD90的表达水平分别为（2.93±0.52）%和（1.30±0.17）%，差异有统计学意义（P〈0.05）。M2＋shCCL2－KYSE450组和M2＋shCCL2－KYSE450＋ASA组细胞中CD90的表达水平分别为（4.07±0.12）%和（4.73±0.38）%，差异无统计学意义（P＝0.17）。结论阿�
ObjectiveTo investigate the effect of tumor-associated macrophages on the stemness of esophageal cancer cells and the potential mechanism of antiproliferative effects of aspirin （ASA）.MethodsThe effects of aspirin on the stemness characteristics of KYSE-450 cells and KYSE-450 cells co-cultured with M2 macrophages （KYSE-450＋ M2） were performed using spheroid formation assay. After treatment with aspirin, the expression of different chemokines, the core pluripotency gene Nanog and the stem cell marker CD90 in different cell groups were determined by real-time quantitative PCR, flow cytometry and Western blot.ResultsThe number of spheres formed in the ASA and KYSE-450＋ M2 cell groups were 7.00±1.23 and 34.33±2.33, respectively, showing statistically significant difference compared with that of control group （14.50±2.33, all P〈0.05）. The number of spheres in KYSE-450＋ M2＋ ASA cell group were 20.67±2.33, which was significantly lower than that of KYSE-450＋ M2 group （P〈0.05）. The expression levels of Nanog gene in control and ASA groups were 1.00 and 0.50±0.10, respectively, and the difference was statistically significant （P〈0.05）. Moreover, the expression of Nanog gene in cells of KYSE-450＋ M2 group and M2＋ KYSE-450＋ ASA group was 1.74±0.13 and 1.43±0.05, showing statistically significant difference （P〈0.05）. When chemokine CCL2 was knocked down, the levels of Nanog gene in M2＋ shCCL2-KYSE450＋ ASA group and M2＋ shCCL2-KYSE450 group were decreased to 1.22±0.11 and 1.17±0.08, respectively, and there was no statistically significant difference between them （P=0.69）. Flow cytometry analyses showed that the expression levels of CD90 in control and ASA cells were （2.93±0.52）% and （1.30±0.17）%, respectively, and the difference was statistically significant （P〈0.05）. Moreover, the expression levels of CD90 in M2＋ shCCL2-KYSE450 cells and M2＋ shCCL2-KYSE450＋ ASA cells were （4.07±0.12）% and （4.73±0.38）%, respectively, showing no
作者
王栋
岳冬丽
王丹
陈新峰
殷向阳
王亚苹
杨黎
张毅
Wang Dong;Yue Dongli;Wang Dan;Chen Xinfeng;Yin Xiangyang;Wang Yaping;Yang Li;Zhang Yi(Biotherapy Center,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,Chin;Department of Oncology,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China,Department of Clinical Medicine,Zhengzhou University,Zhengzhou 450052,China)
出处
《中华肿瘤杂志》
CAS
CSCD
北大核心
2018年第10期744-749,共6页
Chinese Journal of Oncology
基金
河南省医学科技攻关计划重大项目（201501004）
国家青年科学基金（81602599）
