张莹,金爱,王旭东
ZHANG Ying,JIN Ai,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:探讨上皮生长因子(EGF)对人三阴乳腺癌(TNBC)细胞迁移、侵袭能力的影响及CANP-YAP的介导作用。方法:以人源乳腺癌细胞系MDA-MB-231和468细胞为模型细胞,实验分为磷酸盐缓冲液(PBS)组、EGF组、EGF联合钙蛋白酶抑制剂(Cal)组(EGF联合Cal)及EGF联合钙蛋白酶抑制剂Ⅲ(CIⅢ)组(EGF联合CIⅢ),采用伤口愈合实验检测细胞迁移能力、Transwell检测细胞侵袭能力及Western blot法检测YAP蛋白在细胞中的表达水平。结果:伤口愈合实验和Transwell实验结果显示,EGF组MDA-MB-231、468细胞的迁移率和侵袭率均高于PBS组(P<0.05或P<0.01),EGF联合Cal组和EGF联合CIⅢ组中MDA-MB-231、468细胞的迁移率和侵袭率均低于EGF组(P<0.05);Western blot实验结果显示,EGF组MDA-MB-231细胞和MDA-MB-468细胞中YAP蛋白表达均高于PBS组(P<0.05或P<0.01);EGF联合Cal组和EGF联合CIⅢ组中MDA-MB-231、468细胞YAP蛋白表达均低于EGF组(P<0.05或P<0.01)。结论:EGF具有诱导TNBC细胞MDA-MB-231、468细胞迁移和侵袭,Cal和CIⅢ可以阻断EGF的上述生物学效应,EGF可能通过CANP-YAP信号通路促进肿瘤细胞的恶性生物学行为。
Objective: To investigate the effect of epithelial growth factor(EGF) on the migration and invasion of human triple negative breast cancer(TNBC) cells and the mediation of CANP-YAP pathway. Methods: Human breast cancer cell lines MDA-MB-231 and MDA-MB-468 were used as model cells, which were divided into the phosphate buffer(PBS) group, EGF group, EGF combined with calpeptin(Cal) group, and EGF combined with calpain inhibitor(CI) III group. The migration ability of the cells was measured by wound healing test. Transwell was used to detect the invasive ability of the cells, and Western blot was used to detect the expression level of target protein in cells. Results: The migration rate of MDA-MB-231 and-468 cells in EGF group was higher than that in PBS group(P<0.05), and the invasion rate of the two kinds of cells in EGF group was higher than that in PBS group(P<0.05 or P<0.01); The YAP protein expression of the two kinds of cells in EGF group was higher than that in PBS group(P<0.05 or P<0.01); The migration rate of the two kinds of cells in EGF combined with Cal group and EGF combined with CI III group was lower than that in EGF group(P<0.05).The invasion rate of the two kinds of cells in EGF combined Cal group and EGF combined CI III group was lower than that in EGF group(P<0.05), and the expression of YAP protein in EGF combined Cal group and EGF combined CI III group was higher than that in EGF group(P<0.05 or P<0.01). Conclusion: EGF has the ability to induce migration and invasion of model cells. Cal and CI III can block the above biological effects induced by EGF, suggesting that EGF can promote malignant biological behavior of cancer cells through CANP-YAP signaling pathway.

关键词(KeyWords)： 乳腺肿瘤;细胞迁移;上皮生长因子;钙蛋白酶;YES相关蛋白
triple negative breast cancer;cell migration;epithelial growth factor;calpain;YES associated protein

Abstract:

Keywords:

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

作者(Author): 张莹,金爱,王旭东
ZHANG Ying,JIN Ai,WANG Xudong

DOI: 10.19367/j.cnki.1000-2707.2020.01.005

参考文献(References)：

• [1] NAVRATIL J,FABIAN P,PALACOVA M,et al.Triple negative breast cancer[J].Klin Onkol,2015,28(6):405-415.
• [2] 石静,牛立蓉.三阴性乳腺癌转化医学药物领域研究进展[J].实用医学杂志,2018,34(10):1602-1604.
• [3] 黄钱,甘淋.雄激素受体在三阴性乳腺癌中的研究进展[J].实用医学杂志,2019(2):318-321.
• [4] YOON R,STASINOPOULOS I,KIM J H,et al.COX-2 dependent regulation of mechanotransduction in human breast cancer cells[J].Cancer Biol Ther,2015,16(3):430-437.
• [5] YU M,BARDIA A,WITTNER B S,et al.Circulating breast tumor cells exhibit dynamic changes in epithelial andmesenchymal composition[J].Science,2013,339(6119):580-584.
• [6] 王晶晶,彭博,贺荣,等.筋骨草和茯苓配伍合用抑制乳腺癌细胞侵袭转移作用及初步机制研究[J].中国药理学通报,2017,33(4):581-588.
• [7] SCHMIDMAIER R,BAUMANN P.ANTI-ADHESION evolves to a promising therapeutic concept in oncology[J].Current Medicinal Chemistry,2008,15(10):978-990.
• [8] 李勇杰,于庆龙,潘际刚,等.酒精对乳腺癌细胞表皮生长因子受体-钙激活中性蛋白酶通路及细胞迁移的影响[J].中国癌症杂志,2016,26(10):820-825.
• [9] NAVARINI N F,ARAUJO V C,BROWN A L,et al.The EGF signaling pathway influences cell migration andthe secretion of metalloproteinases by myoepithelial cells in pleomorphic adenoma[J].Tumour Biol,2015,36(1):205-211.
• [10]JIANG W,TIAN W,IJAZ M,et al.Inhibition of EGF-induced migration andinvasion by sulfated polysaccharide of Sepiellamaindroni ink via the suppression of EGFR/Akt/p38 MAPK/MMP-2 signaling pathway in KB cells[J].Biomed Pharmacother,2017(11):95-102.
• [11]齐曾鑫,蔡加君,姚瑜,等.钙蛋白酶的研究进展[J].中国临床神经科学,2013,21(4):456-462.
• [12]GUEGUINOU M,HARNOIS T,CROTTES D,et al.SK3/TRPC1/Orai1 complex regulates SOCE-dependent colon cancer cell migration:a novel opportunity to modulate anti-EGFR mAbactionn by the alkyl-lipid Ohmline[J].Oncotarget,2016,7(24):36168-36184.
• [13]GUO L,TENG L.YAP/TAZ for cancer therapy:opportunities and challenges(review) [J].Int J Oncol,2015,46(4):1444-1452.
• [14]ZANCONATO F,BATTILANA G,CORDENONSI M,et al.YAP/TAZ as the rapeutictargets in cancer[J].Curr Opin Pharmacol,2016(8):26-33.
• [15]PICCOLO S,DUPONT S,CORDENONSI M.The biology of YAP/TAZ:hippo signaling and beyond[J].Physiol Rev,2014,94(4):1287-1312.
• [16]ODLE T G.Precision medicine in breast cancer[J].Radiol Technol,2017,88(4):401-421.
• [17]ZHENG Y,WU C X,ZHANG M L.Breast cancer in China’s epidemic situation and disease characteristics[J].China Oncol,2013(8):561-569.
• [18]KIM J,KONG J,CHANG H,et al.EGF induces epithelial-mesenchymal transition through phospho-Smad2/3-Snail signaling pathway in breast cancer cells[J].Oncotarget,2016(51):85021-85032.
• [19]BUGIDE S,DAVID D,NAIR A,et al.Hematopoietic PBX-interacting protein(HPIP) is over expressed in breast infiltrative ductal carcinoma and regulaes cell adhesion and migration through modulation of focal adhesion dynamics[J].Oncogene,2015,32(35):4601-4612.
• [20]SALEHIN D,FROMBERG I,HAUGK C,et al.Immunhistochemical analysis for expression of calpain 1,calpain 2 and calpastatin in ovarian cancer[J].Eur J GynaecolOncol,2011,34(6):628-635.

文章评论(Comment)：