牛上皮细胞中有效表面标记物的筛选
|
摘要
动物上皮细胞容易被多种病毒感染,上皮细胞表面标记物的检测可以作为表征上皮细胞状态的一种手段,但是大多数上皮细胞表面标记物也会在上皮来源的肿瘤细胞中表达。为了选择高效、独特的上皮标记物,采用蛋白质印迹法和间接免疫荧光技术,在牛肾上皮细胞(KE)、舌上皮细胞(TE)、甲状腺上皮细胞(TYE)以及A549细胞和SW480细胞中,检测了AE3(basic cytokeratin)、AE5(CK 3/2p)、CK7、LH1(CK1/10)、EMA(epithelial membrane antigen)、DesmoplakinⅠ/Ⅱ(DSPⅠ/Ⅱ)、E-cadherin和ZO1(zona occludens-1)等8种上皮标记物的表达。结果显示,与阴性对照相比,AE3、AE5、CK7、EMA及DSPⅠ/Ⅱ在上皮细胞和上皮细胞来源的A549、SW480肿瘤细胞中表达良好;E-cadherin仅在上皮细胞中表达;LH1(CK1/10)和ZO1在所有被检测的细胞中都没有表达。结果表明,AE3、AE5、CK7、EMA及DSPⅠ/Ⅱ可以作为有效的上皮细胞及上皮来源的肿瘤细胞标记物,E-cadherin可以作为特异性的上皮细胞标记物。
Animal epithelial cells are susceptible to multiple viral infections,and detection of epithelial cell markers can be used as a mean to characterize the status of epithelial cells,but most epithelial cell surface markers are also expressed in epithelial-derived tumor cells.To select efficient and unique epithelial markers,the researchers used Western blot and indirect immunofluorescence assay to detect the expression of eight epithelial markers,such as AE3(basic cytokeratin),AE5(CK 3/2 p),CK7,LH1(CK 1/10),EMA(epithelial membrane antigen),DesmoplakinⅠ/Ⅱ(DSP Ⅰ/Ⅱ),E-cadherin and ZO1(zona occludens-1)in bovine renal epithelial cells,tongue epithelial cells,thyroid epithelial cells,A549 cells and SW480 cells.The results showed that compared with the negative control,AE3,AE5,CK7,EMA and DSP Ⅰ/Ⅱ were well expressed in the epithelial-derived tumor cells,A549 and SW480,and epithelial cells.E-cadherin was only expressed in epithelial cells.LH1 and ZO1 were not expressed in all tested cells.Therefore,AE3,AE5,CK7,EMA and DSP Ⅰ/Ⅱcan be used as effective cell markers for epithelial cells and epithelial-derived tumor cells,and E-cadherin can be used as specific epithelial cell marker.
Animal epithelial cells are susceptible to multiple viral infections,and detection of epithelial cell markers can be used as a mean to characterize the status of epithelial cells,but most epithelial cell surface markers are also expressed in epithelial-derived tumor cells.To select efficient and unique epithelial markers,the researchers used Western blot and indirect immunofluorescence assay to detect the expression of eight epithelial markers,such as AE3(basic cytokeratin),AE5(CK 3/2 p),CK7,LH1(CK 1/10),EMA(epithelial membrane antigen),DesmoplakinⅠ/Ⅱ(DSP Ⅰ/Ⅱ),E-cadherin and ZO1(zona occludens-1)in bovine renal epithelial cells,tongue epithelial cells,thyroid epithelial cells,A549 cells and SW480 cells.The results showed that compared with the negative control,AE3,AE5,CK7,EMA and DSP Ⅰ/Ⅱ were well expressed in the epithelial-derived tumor cells,A549 and SW480,and epithelial cells.E-cadherin was only expressed in epithelial cells.LH1 and ZO1 were not expressed in all tested cells.Therefore,AE3,AE5,CK7,EMA and DSP Ⅰ/Ⅱcan be used as effective cell markers for epithelial cells and epithelial-derived tumor cells,and E-cadherin can be used as specific epithelial cell marker.
引文
[1]Mellas R E,Leigh N J,Nelson J W,et al.ZO-1,Occludin,and E-cadherin expression and organization in salivary glands with Sjogren's syndrome[J].Acta Histochemica Et Cytochemica Off J Jpn Soci Histochem Cytochem,2014,63(1):45-56.
[2]Sloane J P,Ormerod M G.Distribution of epithelial membrane antigen in normal and neoplastic tissues and its value in diagnostic tumor pathology[J].Cancer,1981,47(7):1786-1795.
[3]Attallah A M,El-Far M,Omran M M,et al.Circulating levels and clinical implications of epithelial membrane antigen and cytokeratin-1in women with breast cancer:can their ratio improve the results?[J].Tumor Biol,2014,35(11):10737-10745.
[4]Al-Abbadi M A,Almasri N M,Al-Quran S,et al.Cytokeratin and epithelial membrane antigen expression in angiosarcomas:An immunohistochemical study of 33cases[J].Arch Pathol Lab Med,2007,131(2):288-292.
[5]Iwata J,Fletcher C D M.Immunohistochemical detection of cytokeratin and epithelial membrane antigen in leiomyosarcoma:A systematic study of 100cases[J].Pathol Inter,2010,50(1):7-14.
[6]Trivino López A,Carlos González I,Yamamoto Y,et al.An immunohistochemical study of epithelial membrane antigen,cytokeratin,and vimentin in papillary thyroid carcinoma:Recognition of lethal and favorable prognostic types[J].Cancer(Philadelphia),1993,72(7):2286-2287.
[7]Rong W,Ying H E,Li L,et al.Comparison of two methods for primary culture of epithelial cells from human bioptic tissue of nasopharyngeal carcinoma[J].J South Med Univ,2010,30(12):2667-2670.
[8]Moll R.Cytokeratins as markers of differentiation.Expression profiles in epithelia and epithelial tumors[J].Veroff Pathol,1993,142:1.
[9]Franke W W,Schiller D L,Moll R,et al.Diversity of cytokeratins:Differentiation specific expression of cytokeratin polypeptides in epithelial cells and tissues[J].J Mol Biol,1981,153(4):933-959.
[10]Barak V,Goike H,Panaretakis K W,et al.Clinical utility of cytokeratins as tumor markers.[J].Clin Biochem,2004,37(7):529-540.
[11]Tot T.Cytokeratins 20and 7as biomarkers:usefulness in discriminating primary from metastatic adenocarcinoma[J].Eur J Cancer,2002,38(6):0-763.
[12]Sherwood E R,Berg L A,Mitchell N J,et al.Differential cytokeratin expression in normal,hyperplastic and malignant epithelial cells from human prostate[J].J Urol,1990,143(1):167-171.
[13]Karantza V.Keratins in health and cancer:more than mere epithelial cell markers[J].Oncogene,2011,30(2):127-138.
[14]Raul U,Sawant S,Dange P,et al.Implications of cytokeratin8/18filament formation in stratified epithelial cells:Induction of transformed phenotype[J].Inter J Cancer,2010,111(5):662-668.
[15]Leroy X,Moukassa D,Copin M C,et al.Utility of cytokeratin7for distinguishing chromophobe renal cell carcinoma from renal oncocytoma[J].Eur Urol,2000,37(4):484-487.
[16]Chang S,Oh M H,Ji S Y,et al.Practical utility of insulin-like growth factor II mRNA-binding protein 3,glucose transporter1,and epithelial membrane antigen for distinguishing malignant mesotheliomas from benign mesothelial proliferations[J].Pathol Inter,2015,64(12):607-612.
[17]Herman H K,Abramowsky C R,Metry D,et al.Identification of bile duct paucity in alagille syndrome:Utilizing CK7and EMA immunohistochemistry as a reliable panel for accurate diagnosis[J].Pediatric Develop Pathol,DOI:2015,15-05-1628-OA.1.
[18]Ylermi S.Tight junctions in lung cancer and lung metastasis:a review[J].Int J Clin Exp Pathol,2012,5(2):126-136.
[19]Jolly M K.E-cadherin represses anchorage-independent growth in sarcomas through both signaling and mechanical mechanisms[J].Mol Cancer Res:MCR,2019.DOI:10.1158/1541-7786.MCR-18-0763.
[20]Zhang X,Wang L,Zhang H,et al.Decreased expression of ZO-1is associated with tumor metastases in liver cancer[J].Oncol Lett,2019,17:1859-1864.
[21]杨举伦,邹红,蔡琳,等.桥粒斑蛋白Ⅱ在各种上皮组织中的表达[J].免疫学杂志,1995(1):40-42.
[22]Vasioukhin V,Bowers E,Bauer C,et al.Desmoplakin is essential in epidermal sheet formation[J].Nature Cell Biol,2001,3(12):1076-1085.
[23]肖春,刘虓,胡火珍.desmoplakin基因在斑马鱼早期胚胎表达的研究[J].四川动物,2010,29(6):965-967.
[24]Abreuvelez A M,Howard M S,Yi H,et al.Patients affected by a new variant of endemic pemphigus foliaceus have autoantibodies colocalizing with MYZAP,p0071,desmoplakins1-2and ARVCF,causing renal damage[J].Clin Exper Dermatol,2018,43(6):692.
[2]Sloane J P,Ormerod M G.Distribution of epithelial membrane antigen in normal and neoplastic tissues and its value in diagnostic tumor pathology[J].Cancer,1981,47(7):1786-1795.
[3]Attallah A M,El-Far M,Omran M M,et al.Circulating levels and clinical implications of epithelial membrane antigen and cytokeratin-1in women with breast cancer:can their ratio improve the results?[J].Tumor Biol,2014,35(11):10737-10745.
[4]Al-Abbadi M A,Almasri N M,Al-Quran S,et al.Cytokeratin and epithelial membrane antigen expression in angiosarcomas:An immunohistochemical study of 33cases[J].Arch Pathol Lab Med,2007,131(2):288-292.
[5]Iwata J,Fletcher C D M.Immunohistochemical detection of cytokeratin and epithelial membrane antigen in leiomyosarcoma:A systematic study of 100cases[J].Pathol Inter,2010,50(1):7-14.
[6]Trivino López A,Carlos González I,Yamamoto Y,et al.An immunohistochemical study of epithelial membrane antigen,cytokeratin,and vimentin in papillary thyroid carcinoma:Recognition of lethal and favorable prognostic types[J].Cancer(Philadelphia),1993,72(7):2286-2287.
[7]Rong W,Ying H E,Li L,et al.Comparison of two methods for primary culture of epithelial cells from human bioptic tissue of nasopharyngeal carcinoma[J].J South Med Univ,2010,30(12):2667-2670.
[8]Moll R.Cytokeratins as markers of differentiation.Expression profiles in epithelia and epithelial tumors[J].Veroff Pathol,1993,142:1.
[9]Franke W W,Schiller D L,Moll R,et al.Diversity of cytokeratins:Differentiation specific expression of cytokeratin polypeptides in epithelial cells and tissues[J].J Mol Biol,1981,153(4):933-959.
[10]Barak V,Goike H,Panaretakis K W,et al.Clinical utility of cytokeratins as tumor markers.[J].Clin Biochem,2004,37(7):529-540.
[11]Tot T.Cytokeratins 20and 7as biomarkers:usefulness in discriminating primary from metastatic adenocarcinoma[J].Eur J Cancer,2002,38(6):0-763.
[12]Sherwood E R,Berg L A,Mitchell N J,et al.Differential cytokeratin expression in normal,hyperplastic and malignant epithelial cells from human prostate[J].J Urol,1990,143(1):167-171.
[13]Karantza V.Keratins in health and cancer:more than mere epithelial cell markers[J].Oncogene,2011,30(2):127-138.
[14]Raul U,Sawant S,Dange P,et al.Implications of cytokeratin8/18filament formation in stratified epithelial cells:Induction of transformed phenotype[J].Inter J Cancer,2010,111(5):662-668.
[15]Leroy X,Moukassa D,Copin M C,et al.Utility of cytokeratin7for distinguishing chromophobe renal cell carcinoma from renal oncocytoma[J].Eur Urol,2000,37(4):484-487.
[16]Chang S,Oh M H,Ji S Y,et al.Practical utility of insulin-like growth factor II mRNA-binding protein 3,glucose transporter1,and epithelial membrane antigen for distinguishing malignant mesotheliomas from benign mesothelial proliferations[J].Pathol Inter,2015,64(12):607-612.
[17]Herman H K,Abramowsky C R,Metry D,et al.Identification of bile duct paucity in alagille syndrome:Utilizing CK7and EMA immunohistochemistry as a reliable panel for accurate diagnosis[J].Pediatric Develop Pathol,DOI:2015,15-05-1628-OA.1.
[18]Ylermi S.Tight junctions in lung cancer and lung metastasis:a review[J].Int J Clin Exp Pathol,2012,5(2):126-136.
[19]Jolly M K.E-cadherin represses anchorage-independent growth in sarcomas through both signaling and mechanical mechanisms[J].Mol Cancer Res:MCR,2019.DOI:10.1158/1541-7786.MCR-18-0763.
[20]Zhang X,Wang L,Zhang H,et al.Decreased expression of ZO-1is associated with tumor metastases in liver cancer[J].Oncol Lett,2019,17:1859-1864.
[21]杨举伦,邹红,蔡琳,等.桥粒斑蛋白Ⅱ在各种上皮组织中的表达[J].免疫学杂志,1995(1):40-42.
[22]Vasioukhin V,Bowers E,Bauer C,et al.Desmoplakin is essential in epidermal sheet formation[J].Nature Cell Biol,2001,3(12):1076-1085.
[23]肖春,刘虓,胡火珍.desmoplakin基因在斑马鱼早期胚胎表达的研究[J].四川动物,2010,29(6):965-967.
[24]Abreuvelez A M,Howard M S,Yi H,et al.Patients affected by a new variant of endemic pemphigus foliaceus have autoantibodies colocalizing with MYZAP,p0071,desmoplakins1-2and ARVCF,causing renal damage[J].Clin Exper Dermatol,2018,43(6):692.