Bosch Patrick P, Stevens Jeff W, Noonan Kenneth J, Buckwalter Joseph A, Midura Ronald J
Department of Orthopaedic Surgery, College of Medicine, The University of Iowa, Iowa City 52242-1181, USA.
Iowa Orthop J. 2002;22:47-54.
Although low-grade cartilage neoplasms typically consist of hyaline-like cartilage, most of them also contain some fibrocartilaginous regions. CD44, a cell surface receptor for hyaluronan, has been identified in cartilage. A family of alternatively spliced mRNA containing the variant 6 (v6) exon sequence of CD44 has been linked to several types of neoplasms. We hypothesized that expressions of v6-containing CD44 species are associated with fibrocartilaginous regions of low-grade cartilage neoplasms. To test this hypothesis we performed reverse transcriptase-polymerase chain reaction (RT-PCR) and immunohistochemical analysis on eight samples: four from normal articular cartilage, one from a synovial chondromatosis, and three chondrosarcomas which were graded as I and I/II. The standard CD44s and a unique v6-containing CD44 species (CD44v6-10) were identified in all tissue samples by RT-PCR analysis. Immunohistochemically, using an antibody that cross-reacted with all CD44 species, CD44 was localized to the cell surface, lacuna wall and intracellular compartment of the chondrocytes in the middle and deep zone of normal cartilage, as well as with cells throughout the neoplastic masses. Utilizing an antibody specific for v6-containing CD44 species, the variant species was identified throughout cells of the middle and deep zone of normal cartilage, and localized selectively to intracellular positions. In neoplastic masses, v6-containing CD44 species were found associated only with cells in the hyaline-like cartilage, but not in the fibrocartilaginous regions. Thus a differential expression of the v6-containing CD44 species in the neoplastic masses containing both hyaline-like cartilage and fibrocartilaginous regions was observed when compared to its homogenous expression in normal hyaline cartilage. An involvement between the lack of the variant CD44 (v6-containing) and altered tissue phenotype (e.g., fibrocartilaginous) is suggested.
尽管低度恶性软骨肿瘤通常由类透明软骨组成,但其中大多数也含有一些纤维软骨区域。CD44是一种透明质酸的细胞表面受体,已在软骨中被鉴定出来。一个包含CD44变体6(v6)外显子序列的可变剪接mRNA家族与几种肿瘤类型有关。我们假设含v6的CD44亚型的表达与低度恶性软骨肿瘤的纤维软骨区域有关。为了验证这一假设,我们对八个样本进行了逆转录聚合酶链反应(RT-PCR)和免疫组织化学分析:四个来自正常关节软骨,一个来自滑膜软骨瘤病,以及三个分级为I级和I/II级的软骨肉瘤。通过RT-PCR分析在所有组织样本中鉴定出了标准的CD44s和一种独特的含v6的CD44亚型(CD44v6-10)。免疫组织化学方面,使用一种能与所有CD44亚型交叉反应的抗体,CD44定位于正常软骨中层和深层软骨细胞的细胞表面、陷窝壁和细胞内区域,以及整个肿瘤块中的细胞。利用一种对含v6的CD44亚型特异的抗体,该变体亚型在正常软骨中层和深层的细胞中均被鉴定出来,并选择性地定位于细胞内位置。在肿瘤块中,含v6的CD44亚型仅与类透明软骨中的细胞相关,而在纤维软骨区域中未发现。因此,与正常透明软骨中其均匀表达相比,在同时含有类透明软骨和纤维软骨区域的肿瘤块中观察到了含v6的CD44亚型的差异表达。提示缺乏变体CD44(含v6)与组织表型改变(如纤维软骨)之间存在关联。