涎腺腺样囊性癌高、低转移细胞系基因表达谱及基质金属蛋白酶表达差异

目的筛选与涎腺腺样囊性癌转移相关的候选基因，并对其中的候选基因进行初步的验证。方法用限制片段差异显示PCR技术（restriction fragments differential display PCR, RFDD-PCR）建立涎腺腺样囊性癌高、低转移细胞株（ACC-M、ACC-2）的表达谱。对两个表达谱的片段进行比较，通过生物信息学的分析，初步筛选出候选基因。用半定量逆转录PCR技术对筛选出的基因进行初步验证。结果RFDD-PCR方法共获得5420个基因片段，其中包含12个基质金属蛋白酶（matrix metalloproteinase，MMP）基因。半定量逆转录PCR方法发现MMP2、MMP7、MMP9、MMP14、MMP15、MMP24在ACC-M和ACC-2中的表达存在明显差异。结论构建了ACC-M和ACC-2细胞株的表达谱，为寻找目的基因奠定了基础。发现MMP2、MMP7、MMP9和MMP15与涎腺腺样囊性癌的发生、发展、转移有关，不同肿瘤细胞的转移能力可能与不同的MMPs家族基因相关。     

Warthin-like tumour of the thyroid gland: RET/PTC expression indicates it is a variant of papillary carcinoma

AIMS: Three cases with features of so-called 'Warthin-like tumour' of the thyroid (WaLTT) are described, in order to evaluate its relationship with papillary carcinoma (PC). METHODS AND RESULTS: We performed an histological and immunohistochemical study with emphasis on RET/PTC expression. The most striking features are represented by marked lymphocytic infiltration in the stalks of papillae and by oxyphilic metaplasia of epithelium, resembling Warthin tumour of the salivary gland. In all cases, we found nuclear features reminiscent of PC. The neoplastic cells were strongly positive for Leu M1 and epithelial membrane antigen (EMA), less for thyroglobulin and negative for calcitonin. The lymphocytic infiltrate was composed of a mixed population of B and T-cells with sparse S100-positive Langerhans cells. An interesting finding was the strong positivity with the antibody against RET/PTC. CONCLUSION: All clinicopathological data along with the presence of the extensive lymphocytic infiltrate could imply a more favourable prognosis. The expression of RET/PTC fusion gene adds support to the hypothesis that this tumour is a variant of PC, probably related to the oncocytic variant of PC.     

Adenoid cystic carcinoma of minor salivary glands. Analysis of 86 cases

Summary 819 salivary gland tumors in surgical pathology files over a 25-year period were reviewed. Among 117 adenoid cystic carcinomas, 86 were located in minor salivary glands and were selected for a clinico-pathological analysis. Complementary histoenzymological investigations and electron microscopic study were performed on specimens from 7 and 13 patients respectively.Adenoid cystic carcinoma occured in older patients (mean age of 54 years) than the other salivary neoplasms. The sex ratio was 1/1. The tumor was located more often in the palate and, to a lesser degree in the buccal floor, tongue or gums.Histologically, epithelial nests contained characteristic cyst-like spaces (cylinders) and 3 varieties of such cylinders were described (mucoid, mucohyalin and hyalin). According to the predominant pattern, 3 types of tumors were shown: basaloïd, cribriform and trabecular. A comparison between histological results and clinical behaviour, available in 67 patients, demonstrated positive correlations. The basaloïd form had always a poor prognosis (numerous early recurrences and metastases, frequent lethal evolution). The cribriform type had an intermediate prognosis, better than basaloïd type and less good than trabecular group (100% of patients still alive at 8 years).Histoenzymological studies revealed high level of acid phosphatase, alkaline phosphatase and leucine aminopeptidase activities round cylindromatous cavities. On the other hand, high oxidative enzyme activities were evenly distributed in all cell types.Ultrastructural findings emphasized the immature characters of epithelial tumor cells. These cells contained numerous ribosomes, but few other organelles. Some more differentiated glandular or epidermoid cells were scattered in neoplastic islands. Rare myoepithelial cells lay in periphery of lobules. Cylinder-like spaces were filled with replicated basal lamellae, mucopolysaccharidic granules and fibrillar structures (microfibrils and periodic collagen fibrils).In the light of these results the histogenesis of this neoplasm was discussed. Like the pleomorphic adenoma, adenoid cystic carcinoma was thought to arise from intercalated ducts. Unable to acquire any high degree of differentiation, this blastomatous tumor had a cellular component almost similar to that shown in intermediate stage of salivary gland embryogenesis.The authors wish to thank M.A. Leost and M. Tacnet for their technical assistance     

The chemokine receptor CCR7 is expressed on epithelium of non-inflamed gastric mucosa, Helicobacter pylori gastritis, gastric carcinoma and its precursor lesions and up-regulated by H. pylori

CCR7 chemokine-receptor expression on tumour cells of gastric carcinoma has been associated with lymph-node metastasis and is thought to play an important role in metastasis. However, so far it is unknown whether CCR7 is newly up-regulated on gastric carcinoma or already expressed in non-neoplastic gastric epithelium. Therefore, epithelial CCR7 expression was investigated in the process of gastric carcinogenesis: non-inflamed mucosa --Helicobacter pylori gastritis -- intestinal metaplasia/dysplasia -- gastric carcinoma. CCR7 was expressed by gastric epithelium in non-inflamed gastric mucosa (n = 5), H. pylori gastritis (n = 17), intestinal metaplasia (n = 10), dysplasia (n = 3) and on tumour cells in 20 of 24 patients with gastric carcinoma (13/14 intestinal-type; 7/10 diffuse-type) as tested by immunohistochemistry. As CCR7 expression by gastric epithelium was significantly stronger in H. pylori gastritis than in non-infected mucosa, the influence of H. pylori on CCR7 receptor expression of gastric epithelial cells was investigated by fluorescence activated cell sorter analysis. H. pylori strains up-regulated the CCR7 chemokine-receptor in CCR7-positive cell lines. No difference in CCR7 up-regulation between cag(+) and cag(-)H. pylori strains was found. Epithelial CCR7 up-regulation by H. pylori may alter the metastatic fate of gastric carcinoma. Additionally, CCR7 expression not only on gastric carcinoma, but also on non-neoplastic gastric epithelium, suggests a novel biological function.     

原发性食管小细胞癌16例临床分析

目的探讨原发性食管小细胞癌的临床特点、治疗及预后。方法回顾分析16例原发性食管小细胞癌患者的临床资料。结果1年总生存率为56．2％，2年总生存率25．0％，3年总生存率12．5％。中位生存期15．7个月。结论对于原发性食管小细胞癌应采用以化疗占主要地位的综合治疗，局限期以手术联合化疗，广泛期以放疗联合化疗为宜。     

NF-κB在人肝细胞肝癌中的表达及与HBV X蛋白的关系

目的：研究核转录因子NF－-κB在人肝细胞肝癌组织中的表达及其与乙型肝炎病毒（HBV ）X蛋白的关系。方法；用免疫组织化学S－P法，检测52例人肝细胞肝癌组织中核转录因子NF－-κB及HBV X蛋白的表达；用脂质体介导的基因转染法将HBV x基因真核表达载体pcDNA3.1-HBX转染入人肝癌细胞系HCC－9204，检测肝癌细胞内核转录因子NF－-κB的表达。结果：52例人肝细胞肝癌组织均有核转录因子NF－-κB的广泛表达，并且在11例HBV X蛋白阳性的肝癌组织，核转录因子NF－-κB位于细胞胞质和胞核，而在41例HBV X蛋白阴性的肝癌组织，核转录因子NF－-κB位于肝癌细胞的胞质。将HBV x基因真核表达载体pcDNA3.1-HBX转染 入人肝癌细胞系HCC－9204，并在稳定表达X蛋白 的肝癌细胞，核转录因子NF－-κB定位于其胞质和胞核，而未进行基因转染的亲体细胞，核转录因子NF－-κB仅定位于细胞质，细胞核无核转录因子NF－-κB的表达。结论：核转录因子NF－-κB在人肝细胞肝癌组织中广泛表达，人肝细胞肝癌中存在着核转录因子NF－-κB的异常激活，并且核转录因子NF－-κB的异常激活与HBV X蛋白有关，X蛋白激活核转录因子NF－-κB， 使其从细胞转位于细胞核，这可能在HBV相关的人原发性肝癌肝癌的发生中起一定作用。     

Transforming growth factor-beta facilitates breast carcinoma metastasis by promoting tumor cell survival

We have shown recently that the hyaluronan receptor, CD44, and matrix metalloproteinase 9 (MMP-9) form a complex on the surface of TA/St mouse mammary carcinoma cells that activates latent transforming growth factor-beta (TGF-β) and is required for tumor invasion. Disruption of the CD44/MMP-9 complex by expression of soluble CD44 results in the loss of tumor invasiveness and abrogates tumor cell survival in host lung parenchyma following intravenous injection into syngeneic mice. To explore the molecular nature of the survival signals derived from the CD44/MMP-9 complex during the development of tumor metastasis, we investigated the possibility that activation of latent TGF-β by the CD44/MMP-9 complex is responsible for tumor cell survival in host lung parenchyma. TA3 cells overexpressing dominant negative soluble CD44 (TA3sCD44), which compromises native CD44 function and the ability of TA3 cells to develop metastases, were transfected with constitutively active or latent TGF-β2 and tested for their ability to form tumors in syngeneic mice. Our results demonstrate that expression of the constitutively active, but not the latent, form of TGF-β2 rescues TA3sCD44 cells from apoptosis during lung colonization. These observations provide evidence that activation of latent TGF-β constitutes an event downstream of CD44-dependent signals that is required for tumor cell survival and metastatic colony formation. The functional axis composed of CD44, MMP-9 and TGF-β may therefore play an important role in the metastatic proclivity of selected tumor types. Abbreviations: ECM – extracellular matrix; HA – hyaluronan; HSPG – heparan sulfate proteoglycan; MMP – matrix metalloproteinase; TGF-β– transforming growth factor β This revised version was published online in July 2006 with corrections to the Cover Date.     

Comparison of anatomical location of squamous cell carcinoma within the oral cavity and oropharynx with the incidence of in vitro hyperdiploidy

The anatomical location of the squamous cell carcinoma (SCCA) within the oral cavity and oropharynx influenced the association of SCCA with the biomarker in vitro hyperdiploidy in human dermal fibroblast cultures (IVH). There was a strong association of IVH with the occurrence of SCCA in the anterior 2/3 of the tongue, floor of the mouth and lower alveolar ridge of the oral cavity and in the base of the tongue and pharyngeal wall of the oropharynx. There was a lower association of SCCA with IVH in the tonsillar region of the oropharynx. IVH showed no association with SCCA located in other anatomical parts of the oral region. The patient group whose diagnosis of SCCA in the anterior 2/3 of the tongue occurred prior to the age of 50 years were invariably IVH-, whereas those diagnosed after the age of 50 years were IVH+, providing evidence for heterogeneity. There was no such correlation of biomarker subgrouping with age of diagnosis demonstrated for SCCA at any other anatomical location within the oral cavity or oropharynx.     

Faster gastric emptying for glucose-polymer and fructose solutions than for glucose in humans

Summary This study examined the rates of gastric emptying for water and 13 different carbohydrate-containing solutions in seven subjects, using conventional gastric intubation techniques. The rates of gastric emptying for water and a 10% glucose-polymer solution were also measured during 90 min of treadmill running at 75% of each subject's maximum oxygen consumption . At rest, 15% glucose-polymer (P) and fructose (F) solutions emptied more rapidly from the stomach and provided a faster rate of carbohydrate delivery than did a 15% glucose (G) solution (p<0.05). The G solutions showed a constant energy delivery rate of 3.3 kcal · min^–1; energy delivery from P and F solutions rose with increasing solution concentrations. The osmolality of the gastric aspirate predicted the rate of gastric emptying for all solutions (p<0.05) but overestimated rates of emptying for 10% and 15% P solutions and underestimated emptying rates for 10% and 15% F solutions. Exercise at 75% decreased the rate of gastric emptying of water but not of 10% P solutions. Thus the different rates of gastric emptying for different carbohydrate-containing solutions were not entirely explained by differences in osmolality. Furthermore, exercise may have different effects on the gastric emptying rates of water and carbohydrate solutions.     

Human telomerase catalytic subunit gene re-expression is an early event in oral carcinogenesis

AIMS: Detection of telomerase catalytic subunit (hTERT) mRNA has been used as a surrogate marker for estimation of telomerase activity. The exact role and timing of telomerase re-activation, a key enzyme implicated in cellular immortalization and transformation, in the multistep process of oral carcinogenesis is still unknown. The aim was to test the hypothesis that (i) quantitative rather than qualitative differences exist in the level of hTERT mRNA expression between normal oral mucosa, different grades of oral epithelial abnormalities and squamous cell carcinomas of the oral cavity, and that (ii) hTERT gene re-expression is an important, probably early event in oral carcinogenesis. METHODS AND RESULTS: The relative quantity of hTERT mRNA was analysed in 45 frozen oral epithelia representing different morphological stages of oral carcinogenesis classified according to the Ljubljana classification and in 37 oral squamous cell carcinomas, using a commercially available LightCycler Telo TAGGG hTERT Quantification kit. hTERT mRNA was not detected in normal or reactive hyperplastic oral epithelia, but was present in 43% of atypical hyperplasias (premalignant lesions), 60% of intraepithelial carcinomas and 68% of oral squamous cell carcinomas. Statistical analysis revealed two groups of oral epithelial changes, with significant differences in the levels of hTERT mRNA expression: 1, normal and reactive hyperplastic oral epithelium, and 2, atypical hyperplasia, intraepithelial carcinomas and squamous cell carcinomas. CONCLUSION: These data suggest that hTERT gene re-expression represents an early event in the multistep process of oral carcinogenesis, already detectable at the stage of precancerous oral epithelial changes. Nevertheless, other genetic aberrations appear to be necessary for progression of oral epithelial abnormalities towards invasive squamous cell carcinoma.