金爱,王宏键,董宇华,王旭东
JIN Ai,WANG Hongjian,DONG Yuhua,WANG Xudong

• 1:贵州医科大学基础医学院生理学教研室

摘要(Abstract)：

目的:探讨钙蛋白酶(Calpain)对17β-雌二醇(E2)诱导人乳腺癌细胞上皮-间质转化(EMT)的影响。方法:以雌激素受体(ER)阳性乳腺癌细胞MCF-7和ER阴性乳腺癌细胞MDA-MB-231细胞为研究模型,分别用二甲基亚砜(DMSO,对照组)、50 nmol/L E2(E2组)及50 nmol/L E2联合10μmol/L Calpeptin(E2+Calpeptin组)处理;采用划痕实验观察各组MCF-7和MDA-MB-231细胞的迁移能力,采用蛋白印迹法检测各组细胞纤连蛋白(FN)和E-钙黏素(E-cadherin)蛋白表达。结果:与对照组比较,E2组MCF-7和MDA-MB-231细胞在24和48 h的迁移率均增加(P <0. 01),细胞中的FN蛋白在24、48和72 h表达均明显上调、E-cadherin蛋白表达明显下调(P <0. 01);与E2组比较,E2+Calpeptin组MCF-7和MDA-MB-231细胞划痕迁移率明显降低,FN表达明显减少、E-cadherin表达明显增加(P <0. 01)。结论:E2可通过Calpain诱导MCF-7和MDA-MB-231乳腺癌细胞的EMT及细胞迁移。
Objective: To investigate the effect of calpain on epithelial-mesenchymal transition( EMT) induced by 17β-estradiol( E2). Methods: Estrogen receptor( ER) positive breast cancer cell line MCF-7( ER +) and ER negative breast cancer cell MDA-MB-231( ER-) were used as model cells. Cells were divided into control group( DMSO),E2 group( 50 nmol/L),and E2 + Calpeptin group( 50 nmol/L E2 + 10 μmol/L Calpeptin); then cell migration ability of MCF-7 and MDA-MB-231 cells was observed by wound healing assay,and the expression of fibronectin( FN) and Ecadherin( E-cadherin) was detected by Western blot. Results: Compared with the control group,the migration of MCF-7 and MDA-MB-231 cells were increased significantly in 24 h and 48 h( P < 0. 01)and the expression of FN was significantly increased and E-cadherin significantly decreased in E2 group at 24 h,48 h and 72 h( P < 0. 01). Compared with the E2 group,cell migration in E2 + Calpeptin group was significantly decreased( P < 0. 01),while the expression of FN was significantly decreased and E-cadherin was significantly increased( P < 0. 01). Conclusion: E2 can induce EMT and cell migration of MCF-7 and MDA-MB-231 breast cancer cells via calpain.

关键词(KeyWords)： 细胞运动;17β-雌二醇;钙蛋白酶;乳腺癌细胞;上皮-间质转化
cell movement;17β-estradiol(E2);calpain;breast cancer cells;epithelial-mesenchymal transition(EMT)

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金资助项目(31660345,31360252)

作者(Author): 金爱,王宏键,董宇华,王旭东
JIN Ai,WANG Hongjian,DONG Yuhua,WANG Xudong

DOI: 10.19367/j.cnki.2096-8388.2020.12.003

参考文献(References)：

• [1]MERINO D,WEBER T S,SERRANO A,et al. Barcoding reveals complex clonal behavior in patient-derived xenografts of metastatic triple negative breast cancer[J].Nature communicationsvol,2019,10(1):766.
• [2]BHOLA N E,JANSEN V M,BAFNA S,et al. Kinomewide functional screen identifies role of PLK1 in hormoneindependent,ER-positive breast cancer[J]. Cancer Research,2019,79(4):876.
• [3]WANG X,LAI Q,HE J,et al. Lnc RNA SNHG6 promotes proliferation,invasion and migration in colorectal cancer cells by activating TGF-β/Smad signaling pathway via targeting UPF1 and inducing EMT via regulation of ZEB1[J]. International Journal of Medical Sciences,2019,16(1):51-59.
• [4]孙志亮，张胜超，李洪利，等．EBP50通过Wnt3a/β-catenin信号通路抑制乳腺癌细胞的侵袭和转移[J]．中国生物化学与分子生物学报，2016(9):1027-1032.
• [5]王旭东，丁姗姗，陈腾祥，等．钙激活中性蛋白酶抑制剂对E2诱导的乳腺癌细胞增殖效应的影响及其意义[J]．贵州医药，2009,33(4):302-304.
• [6]CHEN Y,LI Z,HE Y,et al. Estrogen and pure antiestrogen fulvestrant(ICI 182780)augment cell-matrigel adhesion of MCF-7 breast cancer cells through a novel G protein coupled estrogen receptor(GPR30)-to-calpain signaling axis[J]. Toxicology and Applied Pharmacology,2014,275(2):176-81.
• [7]ZHAO L,LI J,LIU Y,et al. Flotillin1 promotes EMT of human small cell lung cancer via TGF-βsignaling pathway[J]. Cancer Biology Medicine,2018,15(4):400-414.
• [8]PARK S J,KIM JG,KIM N D,et al. Estradiol,TGF-β1and hypoxia promote breast cancer stemness and EMT-mediated breast cancer migration[J]. Oncology Letters,2016; 11(3):1895-1902.
• [9]LIBERTINI S J,ROBINSON B S,DHILLON N K,et al.Cyclin E both regulates and is regulated by calpain 2,a protease associated with metastatic breast cancer phenotype[J]. Cancer Research,2005,65(23):10700-10708.
• [10]于庆龙，张莹，王宏健，等．17β-雌二醇对乳腺癌细胞迁移的影响及CANP-FN通路的介导作用[J]．中国药理学通报，2017,33(10):1371-1376.
• [11]金爱，王宏键，王旭东．表皮生长因子对乳腺癌细胞上皮-间质转化的影响及钙蛋白酶的介导作用[J]．贵州医科大学学报，2019,44(4):412-417.
• [12]LI N,DENG Y,ZHOU L. Global burden of breast cancer and attributable risk factors in 195 countries and territories,from 1990 to 2017:results from the global burden of disease study 2017[J]. J Hematol Oncol,2019,12(1):140-146.
• [13]MERINO D,WEBER T S,SERRANO A,et al. Barcoding reveals complex clonal behavior in patient-derived xenografts of metastatic triple negative breast cancer[J].Nature communications,2019,10:766.
• [14]FLEUROT E,GOUDIN C,HANOUX V,et al. Estrogen receptor-αregulates the expression of syndecan-1 in human breast carcinoma cells[J]. Endocr Relat Cancer,2019,26(6):615-628.
• [15]VYDRA N,JANUS P,TOMA J A,et al. 17β-estradiol activates HSF1 via MAPK signaling in ERα-positive breast cancer cells[J]. Cancers(Basel),2019,11(10):15-33.
• [16]HSU Y L,YEN M C,CHANG W A,et al. CXCL17-derived CD11b(+)Gr-1(+)myeloid-derived suppressor cells contribute to lung metastasis of breast cancer through platelet-derived growth factor-BB[J]. Breast Cancer Research,2019,21(1):23.
• [17]MAIMAITIAILI A,WU D,LIU Z,et al. Erratum to analysis of factors related to non-sentinel lymph node metastasis in 296 sentinel lymph node-positive Chinese breast cancer patients[J]. Cancer biology medicine,2018,15(4):478.
• [18]SHARIAT RAZAVI S M,FORGHANIFARD M M,KORDITAMANDANI D M, et al. MAML1 regulates EMT markers expression through NOTCH-independent pathway in breast cancer cell line MCF7[J]. Biochemical Biophysical Research Communications,2019,510(3):376-382.
• [19]REN Z,YANG T,ZHANG P,et al. SKA2 mediates invasion and metastasis in human breast cancer via EMT[J].Molecular Medicine Reports,2019,19(1):515-523.
• [20]张超，任静文，梁迪，等．OCT4与EMT相关因子在浸润性乳腺癌组织中的表达及其临床意义[J]．中国肿瘤生物治疗杂志，2016,23(4):525-530
• [21]HOU J,WANG X,LI Y,et al. 17beta-estradiol induces both up-regulation and processing of cyclin E in a calpain-dependent manner in MCF-7 breast cancer cells[J].FEBS Letters,2012,586(6):892-96.
• [22]STORR S J,CARRAGHER N O,FRAME M C,et al.The calpain system and cancer[J]. Nature Reviews Cancer,2011,11(5):364-374.

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