Fluge Øystein, Bruland Ove, Akslen Lars A, Lillehaug Johan R, Varhaug Jan E
Department of Molecular Biology, University of Bergen, and Department of Medical Genetics and Molecular Medicine, Haukeland University Hospital, Norway.
Thyroid. 2006 Feb;16(2):161-75. doi: 10.1089/thy.2006.16.161.
We used cDNA microarrays to study gene expression in fresh frozen papillary thyroid carcinoma (PTC) specimens. Seven clinically aggressive carcinomas were included, comprising poorly differentiated PTC and tumors with extensive local invasion or synchronous distant metastases. Ten differentiated (classic) papillary thyroid carcinomas (PTC) and non-neoplastic thyroid tissues were also investigated. TaqMan quantitative reverse transcriptase-polymerase chain reaction (RT-PCR), in situ hybridization, and immunohistochemistry verified the differential gene expression. The B-Raf gene was mutated with a T-->A transversion at nucleotide 1799 (V600E) in 8 of 10 differentiated PTC, and in 4 of 7 aggressive carcinomas. Among genes markedly and equally over-expressed in carcinomas of both the aggressive and classic PtC groups, compared to normal thyroid tissue, were CBP/p300 transactivator (CItED1), fibronectin, growth/differentiation factor 15, potassium inwardly rectifying channel KCNJ2, glutaminyl peptide cyclotransferase, WNT7A, and dipeptidyl peptidase IV. A marked upregulation in carcinomas of P-cadherin mRNA and protein concomitant with E-cadherin downregulation, indicates a possible P-E cadherin "switch" in PTC. The growth factor homologue Nel-like 2, dual specificity phosphatase 5, the serine protease kallikrein 10, and also the tight junction genes claudin 1 and claudin 16, were upregulated in classic PTC but not in aggressive tumors, which may be consistent with altered cell polarity in the dedifferentiated PtC. The aggressive, poorly differentiated PtC group was specifically characterized by marked upregulation of several genes related to cell proliferation such as cell division cycle 2 (CDC2), CDC7, kinesin-like 5, ubiquitin conjugating enzyme E2C, and topoisomerase IIalpha, and by upregulation of genes encoding extracellular matrix proteins such as seprase, extracellular matrix protein 1, and several collagens. These aggressive tumors were also characterized by overexpression of the integrin ligand periostin, and in some biopsies also of osteopontin and of the upstream Rac-regulator dedicator of cytokinesis 10 (DOCK10). These data are interpreted to be consistent with altered cell motility, extracellular matrix remodeling and increased cell proliferation, as important processes in PTC tumor progression.
我们使用cDNA微阵列研究新鲜冷冻甲状腺乳头状癌（PTC）标本中的基因表达。纳入了7例临床侵袭性癌，包括低分化PTC以及具有广泛局部侵袭或同步远处转移的肿瘤。还研究了10例分化型（经典型）甲状腺乳头状癌（PTC）和非肿瘤性甲状腺组织。TaqMan定量逆转录聚合酶链反应（RT-PCR）、原位杂交和免疫组化验证了基因表达差异。在10例分化型PTC中的8例以及7例侵袭性癌中的4例中，B-Raf基因在核苷酸1799处发生了T→A颠换（V600E）突变。与正常甲状腺组织相比，在侵袭性和经典型PTC组的癌中均显著且同等过度表达的基因有CBP/p300反式激活因子（CItED1）、纤连蛋白、生长/分化因子15、内向整流钾通道KCNJ2、谷氨酰胺基肽环化转移酶、WNT7A和二肽基肽酶IV。P-cadherin mRNA和蛋白在癌中显著上调，同时E-cadherin下调，这表明PTC中可能存在P-E钙黏蛋白“转换”。生长因子同源物Nel样2、双特异性磷酸酶5、丝氨酸蛋白酶激肽释放酶10以及紧密连接基因claudin 1和claudin 16在经典型PTC中上调，但在侵袭性肿瘤中未上调，这可能与去分化PTC中细胞极性改变一致。侵袭性、低分化PTC组的特征尤其在于与细胞增殖相关的几个基因显著上调，如细胞分裂周期2（CDC2）、CDC7、驱动蛋白样5、泛素结合酶E2C和拓扑异构酶IIα，以及编码细胞外基质蛋白的基因上调，如分离酶、细胞外基质蛋白1和几种胶原蛋白。这些侵袭性肿瘤的特征还包括整合素配体骨膜蛋白的过表达，在一些活检标本中骨桥蛋白以及细胞分裂上游Rac调节因子细胞分裂素10的 dedicator（DOCK10）也过表达。这些数据被解释为与细胞运动性改变、细胞外基质重塑和细胞增殖增加一致，这些是PTC肿瘤进展中的重要过程。
