分泌型卷曲相关蛋白1通过MAPK/ERK信号通路抑制结直肠癌细胞的迁移侵袭  

SFRP1 inhibits the migration and invasion of colorectal cancer cells through MAPK/ERK signaling pathway

作　　者：雷建卫 汪媛 贺利荣 LEI Jianwei;WANG Yuan;HE Lirong(Oncology Surgery,Baoji Central Hospital,Baoji,Shaanxi 721008,China;Department of Critical Medicine,Baoji Central Hospital,Baoji,Shaanxi 721008,China)

机构地区：[1]宝鸡市中心医院肿瘤外科,陕西宝鸡721008 [2]宝鸡市中心医院重症医学科,陕西宝鸡721008

出　　处：《安徽医药》2022年第4期777-782,I0003,共7页Anhui Medical and Pharmaceutical Journal

摘　　要：目的探讨分泌型卷曲相关蛋白1(SFRP1)对结直肠癌细胞迁移和侵袭的影响机制。方法本研究起止时间2018年11月至2019年6月。运用实时荧光定量反转录聚合酶链反应(qRT-PCR)检测正常结肠上皮细胞HCoEpiC、人结直肠癌细胞HCT8、SW480、RKO中SFRP1的mRNA表达;将pcDNA 3.1组(转染pcDNA 3.1)、pcDNA 3.1-SFRP1组(转染pcDNA 3.1-SFRP1)、pcDNA 3.1-SFRP1+DMSO组[转染pcDNA 3.1-SFRP1,再用二甲基亚砜(DMSO)处理]、pcDNA 3.1-SFRP1+IGF组[转染pcDNA 3.1-SFRP1,再用丝裂原活化蛋白激酶(MAPK)/细胞外信号调节激酶(ERK)信号通路激活剂(IGF1)处理],均用脂质体法转染至HCT8细胞。蛋白质印迹法(Western blotting)检测细胞中SFRP1、波形蛋白(Vimentin)、纤维黏连蛋白(FN)、基质金属蛋白酶9(MMP-9)、N-钙黏蛋白(N-cadherin)、上皮钙黏蛋白(E-cadherin)、MAPK/ERK信号通路关键基因(Ras)、雷帕霉素靶蛋白(mTOR)、细胞外信号调节激酶1/2(ERK1/2)、磷酸化细胞外信号调节激酶1/2(p-ERK1/2)、细胞外信号调节激酶(ERK)的蛋白表达;迁徙实验(Transwell)检测细胞的迁移侵袭。结果与正常结肠上皮细胞HCoEpiC相比,人结直肠癌细胞HCT8、SW480、RKO中SFRP1的表达显著降低[mRNA(1.00±0.06)比(0.36±0.03)、(0.62±0.05)、(0.59±0.05);蛋白(1.00±0.04)比(0.24±0.02)、(0.48±0.04)、(0.47±0.04),均P<0.05]。过表达SFRP1可抑制HCT8细胞的迁移(120±11)比(65±6)和侵袭(98±8)比(47±4),并下调Vimentin(0.96±0.05)比(0.23±0.02)、FN(1.00±0.07)比(0.51±0.04)、MMP-9(0.98±0.08)比(0.35±0.03)、N-cadherin(1.01±0.09)比(0.43±0.04),上调E-cadherin(0.99±0.06)比(3.71±0.27),抑制MAPK/ERK信号通路关键蛋白Ras(0.97±0.07)比(0.34±0.03)、mTOR(1.03±0.07)比(0.42±0.04)、p-ERK1/2(0.98±0.08)比(0.29±0.03)和ERK(1.01±0.06)比(0.31±0.03)的表达。激活MAPK/ERK信号通路可逆转过表达SFRP1对结直肠癌细胞迁移和侵袭的抑制作用。结论SFRP1可抑制结直肠癌细胞的迁移和侵袭,其机制可能与失活MObjective To investigate the mechanism of secretory crimp protein 1(SFRP1)on migration and invasion of colorectal cancer cells.Methods The start and end time of study was from November 2018 to June 2019.Real-time fluorescent quantitative re⁃verse transcription polymerase chain reaction(qRT-PCR)was used to detect the expression of SFRP1 mRNA in normal colonic epitheli⁃al cells HCoEpiC,human colorectal cancer cells HCT8,SW480 and RKO.pcDNA 3.1 group(transfected pcDNA 3.1),pcDNA 3.1-SFRP1 group(transfected pcDNA 3.1-SFRP1),pcDNA 3.1-SFRP1+DMSO[transfected pcDNA3.1-SFRP1 and treated with dimethyl sulfoxide(DMSO)],pcDNA 3.1-SFRP1+IGF(transfected pcDNA 3.1-SFRP1 and treated with MAPK/ERK signaling activator IGF1),all were transfected into HCT8 cells by liposome method.Western blotting(Western blot)was used to detect the proteins expression of SFRP1,Vimentin(Vimentin),fibronectin(FN),matrix metalloproteinase 9(MMP-9),N-cadherin(N-cadherin)and epithelial cadherin(E-cadherin),key genes of mitogen activated protein kinase(MAPK)/extracellular signal-regulated protein kinase(ERK)signaling path⁃way(Ras),rapamycin target protein(mTOR),extracellular signal-regulated kinase 1/2(ERK1/2),phosphorylated extracellular signal-regulated kinase 1/2(p-ERK1/2),extracellular signal-regulated kinase(ERK).The cell migration and invasion were detected by Tran⁃swell chamber.Results Compared with normal colonic epithelial cells HCoEpiC,the expression of SFRP1 in human colorectal cancer cells HCT8,SW480 and RKO were significantly decreased[mRNA(1.00±0.06)vs.0.36±0.03),(0.62±0.05,(0.59±0.05);protein(1.00±0.04)vs.(0.24±0.02),(0.48±0.04),(0.47±0.04)](P<0.05).Overexpression SFRP1 inhibited the migration(120±11)vs.(65±6)and invasion(98±8)vs.(47±4),down-regulated vimentin(0.96±0.05)vs.(0.23±0.02),FN(1.00±0.07)vs.(0.51±0.04),MMP-9(0.98±0.08)vs.(0.35±0.03),N-cadherin(1.01±0.09)vs.(0.43±0.04),up-regulated E-cadherin(0.99±0.06)vs.(3.71±0.27),and inhibited the expression of key proteins of MAPK/ERK signaling pathway including

关 键 词：结直肠肿瘤 细胞外信号调节MAP激酶类 钙黏着糖蛋白类 波形蛋白 分泌型卷曲相关蛋白1 MAPK/ERK信号通路 迁移 侵袭 

分 类 号：R735.34[医药卫生—肿瘤]