摘要
目的探讨CXC趋化因子配体5(CXCL5)通过Wnt/β-连环蛋白(Wnt/β-catenin)信号通路对胃癌细胞小鼠移植瘤生长的影响。方法选取70例胃癌(GC)患者(GC组)和70例健康人群(健康组)为研究对象,检测各组血清CXCL5水平,采用免疫组织化学染色法检测GC组织中CXCL5的阳性表达率,并分析CXCL5表达与临床病理特征的关系。将40只小鼠分为过表达CXCL5组和对照组,每组20只。取小鼠胃癌细胞株MFC,通过稳定过表达CXCL5和转染空载体建立小鼠移植瘤模型,记录各组小鼠移植瘤的生长情况及小鼠存活情况。采用流式细胞术检测小鼠移植瘤组织中CD4^(+)T淋巴细胞、CD8^(+)T淋巴细胞、调节性T细胞(Treg细胞)比例;采用Western blot法检测小鼠移植瘤组织中CXCL5及Wnt/β-catenin信号通路相关蛋白的表达;采用实时荧光定量聚合酶链反应(RT-qPCR)检测各组小鼠移植瘤组织中细胞周期蛋白D1(Cyclin D1)和原癌基因c-Myc(c-Myc)的mRNA表达。结果GC组血清CXCL5水平高于健康组,差异有统计学意义(P<0.05)。GC组织中CXCL5阳性表达率为67.14%(47/70),高于癌旁组织中的CXCL5阳性表达率35.71%(25/70),差异有统计学意义(P<0.05)。GC患者CXCL5阳性表达与阴性表达在肿瘤分化程度、TNM分期及淋巴结转移方面比较,差异有统计学意义(P<0.05)。过表达CXCL5组小鼠存活率低于对照组,差异有统计学意义(P=0.048);随着时间延长,过表达CXCL5组小鼠肿瘤体积大于对照组,差异有统计学意义(P<0.05)。过表达CXCL5组小鼠肿瘤组织中CXCL5、Wnt1、Wnt2、β-catenin蛋白表达均高于对照组小鼠,差异有统计学意义(P<0.05)。过表达CXCL5组小鼠肿瘤组织中CD4^(+)T细胞、CD8^(+)T细胞比例低于对照组,Treg细胞比例高于对照组,差异有统计学意义(P<0.05)。过表达CXCL5组小鼠肿瘤组织中Cyclin D1 mRNA和c-Myc mRNA表达高于对照组,差异有统计学意义(P<0.05)。结论GC患者癌组织和血清中CXCL5表达水平上调,CXCL5异常高表达可能激活Wnt/β-catenin信号通路影响肿瘤免疫活性,促进GC进展。
Objective To investigate the effect of CXC chemokine ligand 5(CXCL5)on the growth of transplanted tumors of gastric cancer(GC)cells in mice via Wnt/β-catenin signaling pathway.Methods A total of 70 GC patients(GC group)and 70 healthy individuals(healthy group)were selected as study subjects.The serum CXCL5 levels in each group were detected,and the positive rate of CXCL5 expression in GC tissues was determined using immunohistochemical staining.The relationship between CXCL5 expression and clinicopathological features was analyzed.Forty mice were divided into overexpressed CXCL5 group and control group,with 20 mice in each group.The mouse gastric cancer cell line MFC was used to establish a mouse transplanted tumor model by stable overexpression of CXCL5 and transfection with an empty vector.The growth of transplanted tumors and the survival conditions of mice in each group were recorded.The proportions of CD4^(+)T lymphocytes,CD8^(+)T lymphocytes,and regulatory T cells(Treg cells)in the transplanted tumor tissues of mice were detected using flow cytometry.The expressions of CXCL5 and Wnt/β-catenin signaling pathway-related proteins in the transplanted tumor tissues of mice were detected using Western blot.The mRNA expressions of cyclin D1 and proto-oncogene c-Myc(c-Myc)in the transplanted tumor tissues of mice in each group were detected using reverse transcription-quantitative real time polymerase chain reaction(RT-qPCR).Results The serum CXCL5 level in the GC group was higher than that in the healthy group,and the difference was statistically significant(P<0.05).The positive rate of CXCL5 expression in GC tissues was 67.14%(47/70),which was higher than 35.71%(25/70)in adjacent tissues,and the difference was statistically significant(P<0.05).There were significant differences in tumor differentiation degree,TNM stage,and lymph node metastasis between GC patients with positive and negative CXCL5 expressions(P<0.05).The survival rate of mice in the overexpressed CXCL5 group was lower than that in the control group,and the difference was statistically significant(P=0.048).As time elapsed,the tumor volume of mice in the overexpressed CXCL5 group was larger than that in the control group,and the difference was statistically significant(P<0.05).The protein expressions of CXCL5,Wnt1,Wnt2,andβ-catenin in the tumor tissues of mice in the overexpressed CXCL5 group were higher than those in the control group,and the differences were statistically significant(P<0.05).The proportions of CD4^(+)T cells and CD8^(+)T cells in the tumor tissues of mice in the overexpressed CXCL5 group were lower than those in the control group,while the proportion of Treg cells was higher than that in the control group,and the differences were statistically significant(P<0.05).The mRNA expressions of Cyclin D1 and c-Myc in the tumor tissues of mice in the overexpressed CXCL5 group were higher than those in the control group,and the differences were statistically significant(P<0.05).Conclusion The expression levels of CXCL5 in cancer tissues and serum of GC patients are upregulated.Abnormally high expression of CXCL5 may activate the Wnt/β-catenin signaling pathway,affect tumor immune activity,and promote the progression of GC.
作者
赵元洋
徐萍
王建华
ZHAO Yuanyang;XU Ping;WANG Jianhua(Yancheng Clinical College of Xuzhou Medical University,Yancheng,Jiangsu,224001;Department of Gastroenterology,Binhai County People's Hospital of Jiangsu Province,Yancheng,Jiangsu,224500;Department of Gastroenterology,Yancheng First People's Hospital of Jiangsu Province,Yancheng,Jiangsu,224001)
出处
《实用临床医药杂志》
2025年第13期27-32,38,共7页
Journal of Clinical Medicine in Practice
基金
国家自然科学基金项目(31700737)。
