摘要
目的探讨Zeste增强子同源物2(EZH2)对乳腺癌细胞增殖和转移能力的影响。方法选取2022年1月至2023年5月河南省人民医院收治切除的203例乳腺癌组织和癌旁组织作为研究对象,采用免疫组织化学分析肿瘤组织EZH2表达水平。采用对照短发卡RNA(shRNA)和EZH2 shRNA慢病毒感染MDA-MB-231细胞,建立对照组和EZH2 KD组细胞,采用细胞计数试剂盒(CCK-8)、5-乙炔基-2’-脱氧尿苷(EdU)染色和体外移植瘤实验分析两组细胞的增殖能力;采用Transwell和蛋白质免疫印迹分析两组细胞的迁移和上皮-间充质转化能力;荧光定量分析EZH2下游基因表达情况。组间计量数据比较采用t检验。结果癌旁组织EZH2蛋白表达阳性率[(30.17±7.84)%]明显低于乳腺癌组织表达阳性率[(75.08±11.16)%],差异有统计学意义(t=33.080,P<0.05)。对照组细胞CCK-8吸光度(A)值(2.01±0.09)明显高于EZH2 KD组(1.44±0.19),差异有统计学意义(t=6.754,P<0.05)。对照组细胞EdU阳性率[(78.30±4.83)%]明显高于EZH2 KD组[(61.4±9.24)%],差异有统计学意义(t=5.694,P<0.05)。对照组细胞体内成瘤体积[(830.32±64.54)mm3]明显高于EZH2 KD组[(642.83±55.71)mm3],差异有统计学意义(t=6.754,P<0.05)。对照组细胞迁移数量[(135.33±11.99)个]明显高于EZH2 KD组[(103.83±8.40)个],差异有统计学意义(t=4.400,P<0.05)。对照组细胞上皮细胞标志物E-钙黏蛋白(E-cadherin)蛋白表达水平(0.74±0.08)明显低于EZH2 KD组(1.44±0.19),差异有统计学意义(t=6.754,P<0.05)。对照组细胞间质细胞标志物N-钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)和Snail蛋白表达水平(1.34±0.07、1.52±0.14、1.17±0.13)明显高于EZH2 KD组(0.87±0.08、0.95±0.10、0.74±0.07),差异有统计学意义(t=11.030、8.270、7.079,P<0.05)。对照组细胞抑癌基因p21和第10号染色体上缺失与张力蛋白同源的磷酸酯酶基因(PTEN)mRNA表达水平(1.16±0.11、1.00±0.09)明显低于EZH2 KD组(1.77±0.13、1.72±0.14),差异有统
Objective To investigate the effect of Zeste enhancer of Zeste homolog 2(EZH2)on the proliferation and metastasis of breast cancer cells.Methods A total of 203 cases of breast cancer tissues and adjacent tissues resected in our hospital were selected as research objects from January 2022 to May 2023.The expression level of EZH2 in tumor tissues was analyzed by immunohistochemistry.MDA-MB-231 cells were infected with control short hairpin RNA(shRNA)and EZH2 shRNA lentivirus,and the control group and EZH2 KD group were established.The proliferation ability of the two groups was analyzed using cell counting Kit-8(CCK-8),5-ethynyl-2’-deoxyuridine(EdU)staining,and in vitro tumor transplantation experiments.The migration and epithelial mesenchymal transition abilities of the two groups were analyzed by Transwell and Western blotting.The downstream gene expression of EZH2 was analyzed by fluorescence quantitation.The comparison of measurement data between groups was conducted using t-test.Results The positive rate of EZH2 protein expression in adjacent tissues[(30.17±7.84)%]was significantly lower than that in breast cancer tissues[(75.08±11.16)%,t=33.080,P<0.05].The CCK-8 absorbance(A)value in the control group(2.01±0.09)was significantly higher than that in the EZH2 KD group(1.44±0.19,t=6.754,P<0.05).The EdU positive rate in the control group[(78.30±4.83)%]was significantly higher than that in the EZH2 KD group[(61.4±9.24)%,t=5.694,P<0.05].The tumor formation volume in the control group[(830.32±64.54)mm3]was significantly higher than that in the EZH2 KD group[(642.83±55.71)mm3,t=6.754,P<0.05].The number of migrating cells in the control group[(135.33±11.99)cells]was significantly greater than that in the EZH2 KD group[(103.83±8.40)cells,t=4.400,P<0.05].The expression level of E-cadherin protein in the control group(0.74±0.08)was significantly lower than that in the EZH2 KD group(1.44±0.19,t=6.754,P<0.05).The expression levels of interstitial cell markers N-cadherin,Vimentin,and Snail protein in the contr
作者
于博凡
申鹏
闫园
Yu Bofan;Shen Peng;Yan Yuan(Department of Breast Surgery,Henan Provincial People’s Hospital,Zhenzhou 450003,China)
出处
《中华实验外科杂志》
CAS
北大核心
2023年第9期1745-1748,共4页
Chinese Journal of Experimental Surgery
