摘要
目的:探讨乳腺导管原位癌浸润发生过程中肌上皮对腺上皮的调控作用机制。方法:随机选取天津医科大学肿瘤医院2008年5月—2010年7月期间收治的157例乳腺导管原位癌患者(高核级患者63例,中核级患者51例,低核级患者43例),并对这些患者的肿瘤组织标本进行上皮间质转化(epithelial-mesenchymal transition,EMT)相关标记物(Snail和ZEB1)的免疫组织化学染色,以探讨肿瘤细胞核级与EMT过程的发生及肌上皮细胞表达状况之间的相关性。为了深入研究乳腺导管原位癌浸润发生过程中肌上皮细胞对腺上皮细胞的调控作用,本文选取了人乳腺肌上皮表型细胞系Hs578Bst和乳腺腺上皮表型细胞系MCF-7,采用Transwell小室共培养模式,构建了乳腺导管原位癌模型。然后,采用Hs578Bst和MCF-7共培养细胞结合外源性TGFβ1和TGFβ1抑制剂,设计实验组、空白对照组、阳性对照组和阴性对照组,4组细胞培养72 h后,分别观察腺上皮细胞MCF-7的形态变化、迁移与增殖能力以及EMT相关基因(Snail和ZEB1)的蛋白及mRNA表达水平的变化。结果:免疫组织化学染色结果显示,在乳腺导管原位癌组织中肿瘤细胞核的异型性与EMT过程的发生及肌上皮细胞的表达呈正相关。通过构建乳腺导管原位癌模型发现,肌上皮细胞Hs578Bst通过刺激TGFβ1表达来促进腺上皮细胞MCF-7发生形态改变。细胞划痕愈合实验和细胞增殖实验的结果显示,肌上皮细胞Hs578Bst可通过刺激TGFβ1表达来促进腺上皮细胞MCF-7发生迁移和增殖。通过蛋白质印迹法和实时荧光定量PCR法发现,肌上皮细胞Hs578Bst可通过刺激TGFβ1表达来上调腺上皮细胞MCF-7中EMT相关基因(Snail和ZEB1)的蛋白及mRNA表达水平。结论:乳腺肌上皮通过分泌/刺激TGFβ1来促进腺上皮发生EMT过程,进而导致乳腺导管原位癌浸润的发生。
Objective:To investigate the regulatory mechanism of myoepithelial cells on glandular epithelial cells during the invasive process of ductal carcinoma in situ of breast.Methods:A total of 157 patients with ductal carcinoma in situ of breast,treated at the Tianjin Medical University Cancer Hospital from May 2008 to July 2010,were randomly selected(including 63 high nuclear grade patients,51 middle nuclear grade patients,and 43 low nuclear grade patients).Immunohistochemical staining for epithelial-mesenchymal transition(EMT)-related markers(Snail and ZEB1)was performed on tumor tissue specimens from these patients to explore the correlation between tumor cell nuclear grade,EMT process,and expression status of myoepithelial cells.To further investigate the regulatory role of myoepithelial cells on glandular epithelial cells during the invasive process of ductal carcinoma in situ of breast,a co-culture model of human myoepithelial cell line Hs578Bst and adenomatous epithelial cell line MCF-7 was established using Transwell chambers.Experimental,blank control,positive control,and negative control groups were designed by combining co-cultured Hs578Bst and MCF-7 cells with exogenous TGFβ1 and TGFβ1 inhibitors.After 72 hours of culture,morphological changes,migration and proliferation capabilities of MCF-7,as well as the changes in protein and mRNA expression levels of EMT-related genes(Snail and ZEB1),were observed in each group.Results:Immunohistochemical staining results demonstrated a positive correlation between tumor nuclear atypia,EMT activation,and myoepithelial cell expression in ductal carcinoma in situ tissues.The model of ductal carcinoma in situ of breast demonstrated that myoepithelial cells Hs578Bst promoted morphological changes in glandular epithelial cells MCF-7 by stimulating TGFβ1 expression.Wound healing and cell proliferation assays revealed that myoepithelial cells Hs578Bst can enhance migration and proliferation of glandular epithelial cells MCF-7 via TGFβ1 activation.Western blot and real-time quantitative PCR confirmed that myoepithelial cells Hs578Bst can upregulate the protein and mRNA expression levels of EMT-related genes(Snail and ZEB1)in glandular epithelial cells MCF-7 by through TGFβ1 stimulation.Conclusion:Breast myoepithelial cells promote EMT in glandular epithelial cells by secreting/stimulating TGFβ1,thereby contributing to the occurrence of invasion in ductal carcinoma in situ of the breast.
作者
杨阳
贾玉棉
李佳珍
王瑾
刘芳芳
郭晓静
YANG Yang;JIA Yumian;LI Jiazhen;WANG Jin;LIU Fangfang;GUO Xiaojing(Tianjin Medical University Cancer Institute&Hospital,National Clinical Research Center for Cancer,Tianjin Clinical Research Center for Cancer,Tianjin Key Laboratory of Cancer Prevention and Therapy,Key Laboratory of Breast Cancer Prevention and Therapy(Ministry of Education),Tianjin 300060,China)
出处
《肿瘤》
2024年第10期1003-1014,共12页
Tumor
基金
国家自然科学基金(81772840)
天津市医学重点学科(专科)建设项目(TJYXZDXK-012A)
关键词
乳腺导管原位癌
腺上皮
肌上皮
上皮-间质转化
Ductal carcinoma in situ of breast
Glandular-epithelium
Myoepithelium
Epithelialmesenchymal transition
