Plasma membrane proteomics of human embryonic stem cells and human embryonal carcinoma cells
Plasma membrane proteomics of human embryonic stem cells and human embryonal carcinoma cells
复制标题
DOI：
10.1021/pr800056j
复制
发表时间：
2008-07-01
影响因子：
4.4
通讯作者：
Krijgsveld, Jeroen
中科院分区：
文献类型：
--
作者：
Dormeyer, Wilma;van Hoof, Dennis;Krijgsveld, Jeroen
人类胚胎干细胞（hESC）在再生医学中具有巨大的意义，因为它们可以无限期地自我更新，并且可以产生任何成体细胞类型。人胚胎癌细胞（hECCs）是在睾丸肿瘤中发现的hESC的恶性对应物。在培养物中获得染色体异常的hESC基本上与hECC无法区分。核型正常的hESC与hECC的直接比较可能导致理解它们的生长控制机制之间的差异，并有助于在不发生生殖细胞癌的情况下实施干细胞的安全治疗用途。虽然已经报道了hECC和hESC的几个比较，但它们的细胞表面蛋白质组在很大程度上是未知的，部分原因是质膜蛋白质组学仍然是一个重大挑战。在这里，我们提出了一个策略，用于识别质膜蛋白，已被优化的应用程序相对较少的干细胞通常可用，并不需要繁琐的细胞分级分离。该方法导致在hESC系HUES-7和hECC系NT 2/D1中分别鉴定出237和219种特异性质膜蛋白。除了已知的干细胞相关细胞表面标志物如ALP、CD 9和CTNNB之外，还鉴定了大量受体、转运蛋白、信号转导蛋白和细胞-细胞粘附蛋白。我们的研究揭示了Hedgehog和Writ通路的几个成员在hESC和hECC中差异表达，包括NPC 1，FZD 2，FZD 6，FZD 7，LRP 6和SEMA 4D，它们在干细胞自我更新和癌症生长中起着关键作用。在染色体12 p上编码的各种蛋白质在睾丸癌中复制，在hECC中被独特地鉴定。其中包括GAPDH、LDHB、YARS 2、CLSTN 3、CSDA、LRP 6、NDUFA 9和NOL 1，已知它们在睾丸癌中上调。不同的HLA分子被揭示在hESC和hECC的表面上，尽管它们的丰度低。结果进行了比较，与基因组和蛋白质组数据集以前报道的，小鼠胚胎干细胞，人内皮细胞和生殖细胞肿瘤。我们的数据提供了HUES-7和NT 2/D1细胞的表面特征，并将正常的hESC与hECC区分开来，有助于解释它们的“良性”与“恶性”性质。
Human embryonic stem cells (hESCs) are of immense interest in regenerative medicine as they can self-renew indefinitely and can give rise to any adult cell type. Human embryonal carcinoma cells (hECCs) are the malignant counterparts of hESCs found in testis tumors. hESCs that have acquired chromosomal abnormalities in culture are essentially indistinguishable from hECC. Direct comparison of karyotypically normal hESCs with hECCs could lead to understanding differences between their mechanisms of growth control and contribute to implementing safe therapeutic use of stem cells without the development of germ cell cancer. While several comparisons of hECCs and hESCs have been reported, their cell surface proteomes are largely unknown, partly because plasma membrane proteomics is still a major challenge. Here, we present a strategy for the identification of plasma membrane proteins that has been optimized for application to the relatively small numbers of stem cells normally available, and that does not require tedious cell fractionation. The method led to the identification of 237 and 219 specific plasma membrane proteins in the hESC line HUES-7 and the hECC line NT2/D1, respectively. In addition to known stemness-associated cell surface markers like ALP, CD9, and CTNNB, a large number of receptors, transporters, signal transducers, and cell-cell adhesion proteins were identified. Our study revealed that several Hedgehog and Writ pathway members are differentially expressed in hESCs and hECCs including NPC1, FZD2, FZD6, FZD7, LRP6, and SEMA4D, which play a pivotal role in stem cell self-renewal and cancer growth. Various proteins encoded on chromosome 12p, duplicated in testicular cancer, were uniquely identified in hECCs. These included GAPDH, LDHB, YARS2, CLSTN3, CSDA, LRP6, NDUFA9, and NOL1, which are known to be upregulated in testicular cancer. Distinct HLA molecules were revealed on the surface of hESCs and hECCs, despite their low abundance. Results were compared with genomic and proteomic data sets reported previously for,mouse ESCs, hECCs, and germ cell tumors. Our data provides a surface signature for HUES-7 and NT2/D1 cells and distinguishes normal hESCs from hECCs, helping explain their 'benign' versus 'malignant' nature.