Sprouty2蛋白在人乳腺癌MCF-7细胞中的表达及增殖情况研究  

Expression and proliferation of Sprouty2 protein in human breast cancer MCF-7 cells

作　　者：莫安薇[1] 杨生辉[1] 黄琰菁[1] MO An-wei;YANG Sheng-hui;HUANG Yan-jing(Department of Oncology,Hainan Provincial People's Hospital,Haikou Hainan 570000,China)

机构地区：[1]海南省人民医院肿瘤内科,海南海口570000

出　　处：《临床和实验医学杂志》2023年第3期242-245,共4页Journal of Clinical and Experimental Medicine

基　　金：海南省自然科学基金面上项目(编号:819MS117);海南省卫生计生行业科研项目(编号:19A200049)。

摘　　要：目的探究Sprouty2蛋白在人乳腺癌MCF-7细胞中的表达及增殖情况。方法回顾性选取2019年1月至2021年12月间就诊于海南省人民医院肿瘤内科的80例乳腺癌患者临床资料,收集乳腺癌组织标本,对Sprouty2蛋白表达的乳腺癌MCF-7细胞进行培养,根据培养方法的不同分为观察组与对照组,观察组应用Sprouty2蛋白特异性抗体孵育,对照组应用DPBS孵育。对比两组Sprouty2蛋白表达情况;利用siRNA技术对MCF-7细胞Sprouty2基因表达进行干预,根据结果分为siRNA干扰组与未干扰组;应用MTT法对细胞增殖活力予以检测,根据划痕试验结果分为沉默组、DMSO对照组;并采用蛋白质印迹法检测MMP-2、MMP-9、MMP-13表达。结果观察组患者MCF-7细胞中Sprouty2蛋白免疫细胞百分数及平均荧光强度为(31.56±0.55)%、3.21±0.16,均明显低于对照组[(37.31±1.32)%、3.29±0.06],差异均有统计学意义(P<0.05)。siRNA干扰组的Sprouty2相对变化量为0.19±0.05,明显低于未干扰组(1.00±0.14),差异有统计学意义(P<0.05)。MTT试验结果显示,经过siRNA干扰,MCF-7细胞活力较对照组显著升高,沉默组细胞迁徙能力明显高于对照组,差异均有统计学意义(P<0.05)。沉默Sprouty2基因的MCF-7细胞中MMP-2、MMP-9、MMP-13表达量为2.53±0.27、0.89±0.12、2.64±0.26,均明显高于对照组(1.26±0.17、0.48±0.11、1.72±0.13),差异均有统计学意义(P<0.05)。结论Sprouty2蛋白表达失调与乳腺癌的发生密切相关,基因下调会提升MCF-7细胞增殖与侵袭能力。Objective To investigate the expression and proliferation of Sprouty2 protein in human breast cancer MCF-7 cells.Methods The clinical data of 80 breast cancer patients who visited the Department of Oncology of Hainan Provincial People’s Hospital from January 2019 to December 2021 were retrospectively selected.breast cancer tissue samples were collected and Sprouty2 protein expressing breast cancer MCF-7 cells were cultured.According to the different culture methods,they were divided into observation group and control group.The observation group was incubated with Sprouty2 protein specific antibody,and the control group was incubated with DPBS.The expression of Sprouty2 protein was compared between the two groups;the expression of Sprouty2 gene in MCF-7 cells was intervened by siRNA technology,and the results were divided into siRNA interference group and non interference group;MTT method was used to detect the cell proliferation activity.According to the scratch test results,the cells were divided into silence group and DMSO control group;the expressions of MMP-2,MMP-9 and MMP-13 were detected by Western blotting.Results Immunofluorescence assay showed that the percentage of Sprouty2 protein immune cells and the average fluorescence intensity in MCF-7 cells in the observation group were(31.56±0.55)%,3.21±0.16,which were significantly lower than those in the control group[(37.31±1.32)%,3.29±0.06],and the differences were statistically significant(P<0.05).The relative change of Sprouty2 in siRNA interference group was 0.19±0.05,which significantly lower than that in non interference group(1.00±0.14),the difference was statistically significant(P<0.05).The results of MTT assay showed that after siRNA interference,the viability of MCF-7 cells was significantly increased compared with that of the control group,and the migration ability of the silencing group was higher than that of the control group,and the differences were statistically significant(P<0.05).The expressions of MMP-2,MMP-9 and MMP-13 in MCF-7 ce

关 键 词：乳腺癌 Sprouty2蛋白 MCF-7细胞 免疫荧光法 siRNA干扰试验 

分 类 号：R737.9[医药卫生—肿瘤]