CXCR6通过WNT/β-catenin通路促进结直肠癌增殖和侵袭  

CXCR6 promotes proliferation and invasion of colorectal cancer through WNT/β-catenin pathway

作　　者：周超熙[1] 王贵英 于滨[1] 王光林[1] 马洪庆 张风宾[2] 李保坤 牛文博 ZHOU Chao-xi;WANG Gui-ying;YU Bin;WANG Guang-lin;MA Hong qing;ZHANG Feng bin;LI Bao-kun;NIU Wen bo(Second Department of Greneral Surgery,The Fourth Hospital of Hebei Medical Unirersity,Shijiazhuang 050019,China;Department of Gastrointestinal Surgery,The Third Hospital of Hebei Medical University,Shijiazhuang 050051,China)

机构地区：[1]河北医科大学第四医院外二科,河北石家庄050019 [2]河北医科大学第四医院消化内科,河北石家庄050019 [3]河北医科大学第三医院胃肠外科,河北石家庄050051

出　　处：《中华肿瘤防治杂志》2021年第10期733-739,共7页Chinese Journal of Cancer Prevention and Treatment

基　　金：国家癌症中心肿瘤科研专项课题(NCC2017A23)。

摘　　要：目的检测CXCR6基因在结直肠癌(CRC)中的表达.分析其通过WNT/Bcatenin通路促进增殖和侵袭对CRC的影响及作用机制。方法收集2017-10-01-2019-09-01于河北医科大学第四医院手术的68例CRC患者(结肠癌48例、直肠癌20例)癌及癌旁组织(≥3cm)标本,应用实时荧光定量聚合酶链反应(qRT-PCR)检测肿瘤组织CXCR6mRNA的表达.分析CXCR6与患者的临床病理特征关系。进一步细胞实验应用蛋白质印迹法检测结肠癌SW620细胞株中CXCR6表达,应用RNA干扰技术构建siRNACXCR6转染SW620细胞株,噻唑蓝(MTT)实验检测细胞活性,Tran-swell小室实验检测侵袭能力.蛋白质印迹法检测Wnt/β-catenin通路关键因子的表达。使用Wnt/B-catenin信号通路激活剂LiCl处理SW620细胞,检测抑制CXCR6表达同时激活Wnt/β-catenin信号通路对细胞活性和侵袭能力的影响。结果CRC患者肿瘤组织CXCR6 mRNA表达水平(3.743±0.861)高于癌旁组织(0.920±0.145),差异有统计学意义,t=26.662.P<0.001。CRC患者临床分期晚(t=-2.190.P=0.032)、分化程度低(=-3.674.P<0.001)和淋巴结转移阳性(t=-4.360.P<0.001)的患者肿瘤组织CXCR6 mRNA表达水平较高。CXCR6在SW620细胞株中呈高表达(F=26.521,P<0.001)。转染后siRNA-CXCR6组细胞活性在48 h(F=5.793,P=0.014)和72 h(F=13.892,P<0.001)降低;细胞侵袭能力下降(F=17.364.P<0.001);Wnt/β-catenin通路关键因子p-GSK3βser9(F=19.944.P=0.002)、核β-catenin(F=45.404,P<0.001)、转录因子4(TCF4,F=26.095,P=0.001)、c-myc(F=11.139.P=0.010)、CyclinDl(F=21.327,P=0.002)和基质金属蛋白酶9(F=20.934,P=0.002)表达均下降。LiCl处理SW620细胞可促进细胞活性和侵袭能力(F值分别为41.025和24.207,均P<0.001)。结论CRC肿瘤组织及SW620细胞系CXCR6高表达,其可能通过调控Wnt/βcatenin通路促进CRC细胞增殖和侵袭。Objective By decting the expression of CXCR6 gene in colorectal cancer(CRC).we analyzed the efet and mechanism of CXCR6 gene promoting proliferation and invasion through W nt/βcatenin pathway on CRC.Methods Real time quantitative PCR(qRT-PCR)was used to detect the mRNA expression of CXCR6 in colorectal cancer patients(48 cases with colon cancer,and 20 cases with rectal cancer)who underwent surgery in the Fourth Hospital of HeBei Medical University from Oetober 1.2017 to September 1.2019 were selected,and the relationship between CXCR6 and clinicopathological features was analyzed.Western blot was used to detect the expression of CXCR6 in colon cancer SW620 cell line,SW620 cell line was transfected with siRNA-CXCR6 by RNA interference technique,MTT assay was used to de-tect cell activity.Transwell chamber assay was used to determine invasion ability.Western blot was used to detect the expression of key factors in the W nt/B catenin pathway.W nt/βcatenin signaling pathway activator LiCl was used to treat SW620 cells to detect the inhibiting effects of CXCR6 expression and simultancously activating Wnt/p-catenin signaling pathway on cell viability and invasion.SPSS 23.0 software was used to process the data by t test and one way analysis of variance.Results The expression of CXCR6 in tumor tissues of patients with colorectal cancer(3.743±0.861)was sig-nificantly higher than that in adjacent tissues(0.920±0.145)(t=26.662,P<0.001).CRC patients with later elinical stage,poorer differentiation.and positive lymph node metastasis presented with higher levels of CXCR6 mRNA expression in tumor tissues(t=-2.190,P=0.032;t=-3.674,P<0.001;t=-4.360,P<0.001).CXCR6 was highly expressed in colon cancer SW620 cells(F=26.521,P<0.001).After transfection,the cell viability of siRNA CXCR6 group was sig-nificantly reduced at 48 hand 72 h(F=5.793,P=0.014;F=13.892,P<0.001),with its cell invasion ability decreasing significantly(F=17.364,P<0.001).Expression of the key factors in Wntβcatenin pathway,including p GSK3pser9,nuclearβcateni

关 键 词：CXCR6 WNT/Β-CATENIN通路 结直肠癌 增殖 侵袭 

分 类 号：R735.34[医药卫生—肿瘤]