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邓宁3327大鼠前列腺癌不同表型亚系寡糖决定簇表达的差异

Differences in expression of oligosaccharide determinants by phenotypically distinct sublines of the Dunning 3327 rat prostate cancer.

作者信息

Abel P D, Foster C S, Tebbutt S, Williams G

机构信息

Department of Surgery, Royal Postgraduate Medical School, Hammersmith Hospital, London, United Kingdom.

出版信息

J Urol. 1990 Sep;144(3):760-5. doi: 10.1016/s0022-5347(17)39587-3.

Abstract

Oligosaccharides expressed by the 3327-H and 3327-MAT LyLu sublines of the Dunning rat prostate cancer model have been compared in formalin-fixed and routinely paraffin-embedded tumour tissues. Binding by lectins of defined specificity has been employed to identify expression of seven oligosaccharide structures by primary and metastatic prostatic carcinoma cells. Neuraminidase digestion was employed to reveal determinants masked by sialic acid. The presence of core Man alpha 1----3(Man alpha 1----6)Man beta 1----4GlcNAc beta 1----4 determinants recognised by Con-A (Canavalia ensiformis) confirmed expression of complex-type glycoconjugates by plasma membrane and cytoplasmic components of the 3327-H tumour but only by cytoplasmic determinants within 3327 MAT LyLu variant tumour-cells. The only other oligosaccharide freely expressed by either tumour-subline was (GlcNAc beta 1----4GlcNAc beta 1----4-)n, recognised by WGA (Triticum vulgaris). Prior to neuraminidase digestion, PNA (Arachis hypogaea) (which identifies Type I oligosaccharides: Gal beta 1----3GalNAc-) bound to pseudoluminal membranes of the 3327-H tumour. However, ECG (Erythrina cristagalli) (which identifies type II oligosaccharides: Gal beta 1----4GlcNAc-) did not bind to this tumour. Unmasked Type I (Gal beta 1----3GalNAc-) and Type II (Gal beta 1----4GlcNAc-) oligosaccharides were not identified in the MAT-LyLu variant. After neuraminidase digestion, PNA-binding was identified along pseudoluminal plasma membranes within 3327-H tumours but only within the cytoplasm of 3327-MAT LyLu primary and metastatic tumour cells. Following neuraminidase digestion, ECG-binding was observed along pseudoluminal plasma membranes of 3327-H tumours and heterogeneously within the cytoplasm of primary, but not metastatic 3327-MAT LyLu tumours. Terminal alpha/beta GalNAc- residues recognised by SBA (Glycine max) were not freely expressed by either subline. These structures were readily detected along luminal membranes of 3327-H cells and weakly detected within the cytoplasm of primary but not metastatic MAT 3327-LyLu tumour cells following neuraminidase digestion. Fucosylated Type II structures Fuc alpha 1----2Gal(GalNAc)-), recognised by UEA-1 (Ulex europaeus-1) and GalNAc alpha 1----3GalNAc- structures recognised by DBF (Dolichos biflorus) were not identified as a component of either tumour subline. The different patterns of oligosaccharide expression, identified by lectin-binding, clearly differentiated between the two tumour sublines and distinguished them from normal prostatic epithelium. The Dunning 3327 rat prostatic cancer sublines offer a useful model with which to examine the relationship between cell-surface oligosaccharide structures and phenotypic variants within a defined tumour-cell population.(ABSTRACT TRUNCATED AT 400 WORDS)

摘要

在福尔马林固定、常规石蜡包埋的肿瘤组织中,对邓宁大鼠前列腺癌模型的3327 - H和3327 - MAT LyLu亚系所表达的寡糖进行了比较。利用具有特定特异性的凝集素结合来鉴定原发性和转移性前列腺癌细胞中七种寡糖结构的表达。采用神经氨酸酶消化来揭示被唾液酸掩盖的决定簇。刀豆球蛋白A(Con - A,Canavalia ensiformis)识别的核心甘露糖α1→3(甘露糖α1→6)甘露糖β1→4GlcNAcβ1→4决定簇的存在,证实了3327 - H肿瘤的质膜和细胞质成分表达复合型糖缀合物,但仅在3327 MAT LyLu变异肿瘤细胞的细胞质决定簇中表达。两个肿瘤亚系唯一自由表达的其他寡糖是被小麦胚芽凝集素(WGA,Triticum vulgaris)识别的(GlcNAcβ1→4GlcNAcβ1→4 -)n。在神经氨酸酶消化之前,花生凝集素(PNA,Arachis hypogaea)(识别I型寡糖:Galβ1→3GalNAc -)与3327 - H肿瘤的假管腔膜结合。然而,刺桐凝集素(ECG,Erythrina cristagalli)(识别II型寡糖:Galβ1→4GlcNAc -)不与该肿瘤结合。在MAT - LyLu变异体中未鉴定出未被掩盖的I型(Galβ1→3GalNAc -)和II型(Galβ1→4GlcNAc -)寡糖。神经氨酸酶消化后,在3327 - H肿瘤的假管腔质膜上发现了PNA结合,但仅在3327 - MAT LyLu原发性和转移性肿瘤细胞的细胞质中发现。神经氨酸酶消化后,在3327 - H肿瘤的假管腔质膜上观察到ECG结合,在原发性3327 - MAT LyLu肿瘤细胞的细胞质中呈异质性结合,但在转移性肿瘤细胞中未观察到。大豆凝集素(SBA,Glycine max)识别的末端α/βGalNAc - 残基在两个亚系中均未自由表达。神经氨酸酶消化后,这些结构在3327 - H细胞的管腔膜上很容易检测到,在原发性3327 - MAT LyLu肿瘤细胞的细胞质中检测到较弱信号,但在转移性肿瘤细胞中未检测到。未鉴定出被欧洲荆豆凝集素 - 1(UEA - 1,Ulex europaeus - 1)识别的岩藻糖基化II型结构Fucα1→2Gal(GalNAc) - 和被双花扁豆凝集素(DBF,Dolichos biflorus)识别的GalNAcα1→3GalNAc - 结构作为任何一个肿瘤亚系的成分。通过凝集素结合鉴定出的不同寡糖表达模式,清楚地区分了两个肿瘤亚系,并将它们与正常前列腺上皮区分开来。邓宁3327大鼠前列腺癌亚系提供了一个有用的模型,可用于研究特定肿瘤细胞群体中细胞表面寡糖结构与表型变异之间的关系。(摘要截短于400字)

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