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Invited Review Characterisation of Thyroid Medullary Carcinoma TT Cell

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Invited Review Characterisation of Thyroid Medullary Carcinoma TT Cell Histol Histopathol (1997) 12: 283-289 Histology and 001: 10.14670/HH-12.283 Histopathology http://www.hh.um.es From Cell Biology to Tissue Engineering Invited Review Characterisation of thyroid medullary carcinoma TT cell line M. Zabel1,2 and J. Grzeszkowiak1 1Department of Histology and Embryology, Medical Academy, Poznan and 2Department of Histology and Embryology, Medical Academy, Wroclaw, Poland Summary. IT cell line is the best known stabilized cell the parafollicular cells, together with the demonstration line derived from the human medullary thyroid that they are not invariably parafollicular (Cavalheira carcinoma. The ultrastructural characteristics of these and Pearse, 1967) or confined to the thyroid gland cells include well developed rough endoplasmic (Carvalheira and Pearse, 1967), and that they are derived reticulum, a prominent Golgi apparatus and a (penultimately at least), from the ultimobranchial body considerable number of secretory granules. Numerous (Pearse and Carvalheira, 1967) led to the proposal by hormones were immunocytochemically demonstrated in Pearse (1966) that they should be called C cells (C for IT cells of which calcitonin and calcitonin gene-related calcitonin). Therefore, the terms C cells and para­ peptide (CGRP) are the products of the same gene but an follicular cells are in use to describe the same type of alternative RNA processing. TT cells were found to thyroid gland cells, which produce calcitonin. In the produce some other hormones as well, namely ACTH, human thyroid gland, C cells are normally restricted to neurotensin, enkephalin, PTHrP, gastrin-releasing the posterior part of the lateral lobes. peptide (GRP), serotonin but also functional proteins of It is form the parafollicular cells that the medullary the chromogranin group, synaptophysin, NSE, calbindin thyroid carcinoma (MTC) arises. Prior to 1959 it was not and tyrosine hydroxylase. Some marker proteins have distinguished as a tumor separate from anaplastic been detected in the cytosol (CEA) and in the cyto­ carcinoma. Overall, it accounts for about 10% of all skeleton (alpha-tubulin, cytokeratin). The influence of thyroid cancers and occurs in a younger age group. An numerous factors on the secretory activity of these cells estimated 25% of all MTC are familial and are has been demonstrated so far, including effects of 1,25- associated with mUltiple endocrine neoplasia syndromes dihydroxycholecalciferol, glucocorticoids, sex steroids, (MEN). Data from the German MTC Registry indicate cAMP, gastrin-releasing peptide, sodium butyrate, that 16.6% MTC are associated with MEN 2A (Sipple's phorbol esters, ionomycin and forskolin. The syndrome) accompanied by pheochromocytoma and investigators performed on the IT cell line demonstrate abnormalities of the parathyroid. 5.3% MTC are that this is the most reliable model system for the human associated with FMTC (familial MTC alone), and 2.7% parafollicular cells developed so far, in comparison to with MEN 2B characterised by MTC, pheochromo­ other cell lines derived from the medullary carcinoma of cytoma and ocular and oral neuromas with gastro­ the thyroid. intestinal ganglioneuromatosis. The syndrome is inherited as an autosomal dominant trait. 75% of MTCs Key words: IT cell line, Thyroid medullary carcinoma, are nonfamilial. This proportion, however, is likely to Hormones, Functional proteins, Cell culture decrease as a result of detailed genetic analyses of patients with apparent sporadic tumors (DeLellis, 1995). A variety of somatic abnormalities are mostly associated Introduction with the effects of the secretory products of MTC on many tissues and organs (Thomson, 1981; Kohler, The designation «parafollicular» was first given to 1986). The MTC cells were found to produce calcitonin the specialized and supposedly endocrine, nonfollicular (CT), considered as a marker of this neoplasm. CT and cells of the mammalian thyroid gland by Nonidez other substances typical for parafollicular cells (Gagel et (1932). The successful demonstration by immuno­ aI., 1980; Bose et aI., 1992) are val uable in early fluorescence (Bussolati and Pearse, 1967) of calcitonin in diagnosis of MTC especially after pentagastrin stimulation in the screening of potentially affected Offprint requests to: Professor M. Zabel, Medical Academy, ul. members of MEN 2 families (Raue and Grauer, 1994). Swiecickiego 6, PL·60-781 , Poznan, Poland Also, some substances non typical for parafollicular 284 Characterisation of TT eel/line cell s, such as ACTH, neurotensin, enkephalin, PTHrP 1992). and serotonin (Zabel, 1984; Oosterom et aI. , 1986; In the cell s the presence of proteins associated with Zeytin and DeLellis, 1987; Zeytin et aI. , 1987; Ikeda et secretory granules has also been demonstrated. T he a I. , 19 88; Bida rd et aI. , 1993) are in use fo r the protein s include chromogranin A, SP-I and synapto­ screening. physin (Murray et aI. , 1988; Zabel et a\. , 1995). A very As model systems of the C cell s various cell lines important finding involved the detecti on of NSE in IT derived from the MTC have been developed. The first cell s, since the quantity of enolase is known to change CT-secreting WAG/Rij transplantable rat MTC (rMTC) with vari ations in CT secreti ons (Schafer and Zabel, was described by Boorman et al. (1974). They showed 1983; Ka med a, 1985 ; Z a be l a nd Sc hafer, 1988). that the transpl anted neoplasm maintained some of the Intensity of NSE cellular staining has been noted to be morphological and functional characteri sti cs of the inversely related to CT content of the cells and directl y ori gin al tumor (Zeytinoglu , et aI. , 1980). Another well related to CT level in the medium. Therefore, NSE may known stabilized cell line derived from the rMTC is the be used to evalu ate the functi onal statu s of TT cells 44-2C cell line (Zeytin and DeLellis, 1987). The cell s (Zabel et aI. , L99 5) (Fig. 2). were shown to synthesise and secrete neurotensin , CT From amongst the enzymes participating in biogenic and somatostatin (Zeytin et aI. , 1987). The murine CA- amine fo rmation (Zabel, 1985), th yrosin hydroxylase has 77 cell line was also successfull y used as a model system been demonstrated in IT cell s (Zabel et aI. , 1995). of the para fo llicular cell s (Muszynski et aI. , 1983). In the The diagnostic signi ficance of the CEA and cyto­ cell s, expression of the CT/CGRP and cholecystokinin skeleton proteins has been confirmed by immunocyto­ genes has been demonstrated (Odum and Rehfe ld , 1990; chemical studies on medullary carcinomas (Raue, 1985; Colli gnon et aI. , 1992). Miettinen, 1987). The best known stabilized cell line deri ved from As fa r as the locali zati on of secre tory products human MTC is the TT cell line. It was developed by within IT is concerned, the secretory granules contain Leong et al. in 198 1 (subcultures 24 to 30) (Leong et aI. , CT, CGRP, somatostatin, neurotensin , met-enkephalin , 198 1) . It is very importa nt to investi g ate the le u-enke phalin , GRP, parath yroid hormone-re lated characteristics of these cells as they are the most reli able protein , functional proteins of the chromogranin group mode l syste m of the huma n pa rafo llic ul a r cell s and synaptophysin while in the cytosol NS E, calbindin developed so far. and tyrosine hydroxy lase can b e fo und . Some marker When in vesti gating the characteristi cs of cultured proteins have been detected in the cytosol (CEA) and in e ndocrine cell s it is impo rta nt to re me mbe r tha t the cytoskeleton (alpha-tubulin, cytokeratin) (Zabel et neoplastic endocrine cells, particul arl y when in culture, aI. , 1995). may decisively alter the expression of several proteins and thei r hormone expression, as indicated by numerous The structure and expression of CTICGRP gene studies (Chaiwun et aI. , 1994). The structure and expression of the CT gene were of Morphology and immunocytochemistry of TT cells special interest as much of it is known to yield two di stinct mRNAs, whi ch provide templates for production IT cell s exhibit lower number of secretory granules of either CT or CGRP (Amara et at. , 1982; Nelkin et aI. , th an in normal thyroid parafollicul ar cell s (Zabel and 1984; Edbrooke et a\. , 1985; Jonas et a\. , 1985; Sabate et Sc ha fe r, 1988; Za be l e t a I. , 1994). S ome othe r aI. , 1985; Steenbergh et aI. , 1986; Emerson et aI. , 1989). ultrastructural characteri sti cs of IT cell s include well ­ This has been worked out in detail using bi ochemi cal developed rough endopl asmic reticulum present within a techniques (Rosenfeld et aI. , 1984; Jonas et aI. , 1985; restric ted space in the pe rinuc lear regio n a nd a Steenbergh et aI. , 1986). It has been demonstrated that prominent Golgi apparatus (Zabel et aI. , 1994). this gene located on the short arm of chromosome II IT cell s were fo und to produce numerous hormones consists of six exons, of which exons I, 2 and 3 are as well as some func ti o nal prote ins a nd m a rke rs present in each of the mRNAs. Moreover, CT mRNA although in ma n a nd in most ma mmali an species, contain s sequences complementary to exon 4 while inc luding rat, the f irst immunocytochemicall y CGRP mRNA contains sequences fo r exons 5 and 6 demonstrated hormones were CT, calcitonin gene-related (Amara et aI. , 1982; Jonas et aI. , 1985). The expression peptide (CGRP), somatostatin, gastrin-releasing peptide of the CT gene is frequently quoted as an example of (GRP) and ACTH (Gagel et aI. , 1986; Oosterom et a\. , alternate RNA processing. Thus, the CT gene yield s a 1986; Cote et aI.
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