Franzén B, Linder S, Alaiya A A, Eriksson E, Uruy K, Hirano T, Okuzawa K, Auer G
Unit of Cell and Molecular Analysis, Karolinska Institute and Hospital, Stockholm, Sweden.
Br J Cancer. 1996 Nov;74(10):1632-8. doi: 10.1038/bjc.1996.600.
Malignant progression of tumour cells is caused by the accumulation of genetic defects, which when combined will generate a large phenotypic diversity. Simultaneous quantitation of a large number of gene products in tumour cells is desirable, but difficult to achieve. We have here quantitated the levels of a number of abundant polypeptides in human breast carcinoma cells using two-dimensional gel electrophoresis (2-DE; PDQUEST). For this purpose, tumour cells were prepared from the tissue of 17 breast carcinomas. Fibroadenoma tissue was used as reference for benign cells. An increase of the spot density of the PCNA polypeptide was observed in rapidly proliferating tumour cells, confirming the validity of the procedures used. In the set of 24 polypeptide spots with known identity, decreases in cytokeratin and tropomyosin levels were observed. The levels of all cytokeratin forms resolved (CK7, CK8, CK15 and CK18) were significantly lower in carcinomas than in fibroadenomas. The levels of tropomyosin 2 and 3 were lower in carcinomas than in fibroadenomas. In contrast, the levels of some members of the stress protein family (pHSP60, HSP90 and calreticulin) were higher in carcinomas. Furthermore, changes in the expression of lactate dehydrogenase and GT-pi, but not in nm23, were observed. We conclude that simultaneous analysis of multiple polypeptides in human carcinomas can be achieved by 2-DE and may be useful in prognostic studies, and that malignant progression of breast carcinomas results in the decreased expression of cytokeratin polypeptides. This phenomenon must be considered in studies where cytokeratins are used as markers to identify the epithelial cell compartment in breast carcinomas.
肿瘤细胞的恶性进展是由遗传缺陷的积累引起的，这些缺陷结合在一起会产生很大的表型多样性。同时定量肿瘤细胞中大量的基因产物是很有必要的，但很难实现。我们在此使用二维凝胶电泳（2-DE；PDQUEST）定量了人乳腺癌细胞中多种丰富多肽的水平。为此，从17例乳腺癌组织中制备肿瘤细胞。纤维腺瘤组织用作良性细胞的对照。在快速增殖的肿瘤细胞中观察到增殖细胞核抗原（PCNA）多肽的斑点密度增加，证实了所用方法的有效性。在一组已知身份的24个多肽斑点中，观察到细胞角蛋白和原肌球蛋白水平降低。所有分辨出的细胞角蛋白形式（CK7、CK8、CK15和CK18）在癌组织中的水平均显著低于纤维腺瘤。原肌球蛋白2和3在癌组织中的水平低于纤维腺瘤。相反，应激蛋白家族的一些成员（pHSP60、HSP90和钙网蛋白）在癌组织中的水平较高。此外，观察到乳酸脱氢酶和GT-pi表达的变化，但nm23没有变化。我们得出结论，通过二维凝胶电泳可以实现对人癌组织中多种多肽的同时分析，这可能在预后研究中有用，并且乳腺癌的恶性进展导致细胞角蛋白多肽表达降低。在将细胞角蛋白用作识别乳腺癌上皮细胞成分的标志物的研究中，必须考虑这一现象。
