de Nigris F, Visconti R, Cerutti J, Califano D, Mineo A, Santoro M, Santelli G, Fusco A
Servizio di Oncologia Sperimentale E Istituto per lo Studio e la Cura dei Tumori Fondazione G. Pascale, Naples, Italy.
Cancer Res. 1998 Oct 15;58(20):4745-51.
A subtractive library screening was performed to identify changes in gene expression that occur during the process of neoplastic transformation of thyroid cells. A cDNA library was constructed from a human thyroid papillary carcinoma cell line (NPA) subtracted with cDNAs from normal thyroid cells (HTC 2). The differential screening of this library lead to the isolation of 39 cDNA clones; six of them showed homology with a recently isolated gene, named HIP, that codes for a protein belonging to a novel class of heparin/heparan sulfate-binding proteins. Northern blot analysis revealed HIP gene overexpression in all of the human thyroid carcinoma cell lines analyzed, as compared to the HTC 2 cells. HIP expression was particularly abundant in the anaplastic carcinoma-derived cell lines. The analysis of surgically removed thyroid tumors showed overexpression of HIP gene in all of the carcinomas, independent of the histotype, although the largest increase in HIP expression was observed in the undifferentiated forms. In contrast, none of the benign adenomas or normal thyroid tissues showed HIP overexpression. To establish the role of HIP overexpression in cell transformation, the NPA cell line was transfected with an eukaryotic expression vector carrying the HIP gene in the antisense orientation. Stable transfectants expressed reduced HIP mRNA levels and showed morphological changes, such as becoming spindle-shaped and growing scattered. The growth rate of the antisense clones was greatly reduced compared to the NPA cells transfected with the backbone vector. Taken together, these results indicate that HIP gene overexpression is associated with thyroid carcinogenesis and strongly suggest its involvement in thyroid cell growth regulation.
进行了消减文库筛选，以鉴定甲状腺细胞肿瘤转化过程中发生的基因表达变化。从人甲状腺乳头状癌细胞系（NPA）构建了一个cDNA文库，该文库用来自正常甲状腺细胞（HTC 2）的cDNA进行了消减。对该文库的差异筛选导致分离出39个cDNA克隆；其中6个与最近分离的名为HIP的基因显示出同源性，该基因编码一种属于新型肝素/硫酸乙酰肝素结合蛋白类别的蛋白质。Northern印迹分析显示，与HTC 2细胞相比，在所分析的所有人甲状腺癌细胞系中HIP基因均过表达。HIP表达在间变性癌衍生的细胞系中特别丰富。对手术切除的甲状腺肿瘤的分析表明所有癌中HIP基因均过表达，与组织学类型无关，尽管在未分化形式中观察到HIP表达增加最大。相反，良性腺瘤或正常甲状腺组织均未显示HIP过表达。为了确定HIP过表达在细胞转化中的作用，用携带反义方向HIP基因的真核表达载体转染NPA细胞系。稳定转染子表达降低的HIP mRNA水平，并显示出形态学变化，如变成纺锤形并分散生长。与用空载体转染的NPA细胞相比，反义克隆的生长速率大大降低。综上所述，这些结果表明HIP基因过表达与甲状腺癌发生有关，并强烈提示其参与甲状腺细胞生长调节。
