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Similarity of the product of the Drosophila neurogenic gene big brain to transmembrane channel proteins.

Cells in the neurogenic region of Drosophila embryos are initially bipotential; they can become either neuroblasts or epidermoblasts. Cell-cell interaction seems to play an important part in this developmental decision, which involves the function of a group of genes (the neurogenic genes). Loss-of-function mutations in any of the neurogenic genes result in nervous system hyperplasia and epidermal hypoplasia. Of the six known zygotic neurogenic genes, big brain (bib) is unique in several aspects. Most notably, all the other known neurogenic genes seem to fit into a cascade defined by genetic interactions, whereas bib does not show any detectable interaction with them. To understand how bib functions, we have now cloned the bib genomic and complementary DNAs. The predicted bib product shows significant sequence similarity to a family of transmembrane proteins, some of which form channels permeable to small molecules. Together with genetic studies, our results indicate that the bib product may mediate intercellular communication in a pathway separate from the one involving the products of the other neurogenic genes.

Amino Acid Sequence↗

Recent advances in glycomics and glycogenetics.

As the human genome sequence is nearly deciphered, it is important to turn the attention to the physiological functions of the genes. Thus, the study of the gene products, the proteins, is the next big challenge. The proteins, however, are not the final gene products in many cases. It has been shown that carbohydrates participate in post-translational modifications and in many other functional regulations, hence the study of the glycome, the entire collection of carbohydrates is essential in order to determine the functions of all genes, and will greatly enhance the field of chemical genetics. Known biological function / targets of carbohydrates and combinatorial synthesis & structural analysis of natural / non-natural carbohydrates are surveyed in this review. Methods to search for new biological targets that include carbohydrate mimetics and carbohydrate scaffolds along with chip technology, are also presented.

Carbohydrate Metabolism↗

Gene therapy of orthotopic hepatocellular carcinoma in rats using adenovirus coding for interleukin 12.

The use of gene therapy to enhance antitumor immunity has emerged as a promising procedure to fight cancer. In this study we have tested the ability of an adenovirus carrying interleukin 12 (IL-12) gene (AdCMVIL-12) to eliminate tumoral lesions in 3 animal models of orthotopic hepatocellular carcinoma (HCC). Intratumoral injection of AdCMVIL-12 in animals with a single big tumor nodule implanted in the liver resulted in significant inhibition of tumor growth in a dose-dependent manner. Fifty percent of animals that received a dose of 5 x 10(9) plaque-forming units, showed complete regression of the tumor 2 weeks after treatment. In animals with 2 independent tumor nodules in the left liver lobe, injection in only one of them of 5 x 10(9) pfu AdCMVIL-12 induced, 15 days after therapy, complete regression of 50% of treated tumors and also of 50% of untreated lesions, with 60% long-term survival. Rats that were tumor free after therapy with AdCMVIL-12 showed protection against tumor rechallenge. A group of rats received the carcinogen diethylnitrosamine and developed multiple hepatic dysplasic nodules of 1 to 5 mm in diameter. These animals were treated by intrahepatic artery injection of either AdCMVIL-12 (5 x 10(9) pfu) or control vector. In this model AdCMVIL-12 induced complete tumor regression in 20% of treated rats and inhibited tumor growth in 60% of cases with an increase in rat survival. Activation of natural killer (NK) cells and inhibition of angiogenesis were found to be antitumor mechanisms set in motion by AdCMVIL-12. Our data indicate that experimental HCC can be efficiently treated by intratumoral or intravascular injection of adenovirus expressing IL-12.

Adenoviridae↗

Heterodimerization of endothelin-converting enzyme-1 isoforms regulates the subcellular distribution of this metalloprotease.

Endothelin-converting enzyme (ECE) is a membrane metalloprotease that generates endothelin from its direct precursor big endothelin. Four isoforms of ECE-1 are produced from a single gene through the use of alternate promoters. These isoforms share the same extracellular catalytic domain and contain unique cytosolic tails, which results in their specific subcellular targeting. We investigated the distribution of ECE-1 isoforms in transfected AtT-20 neuroendocrine cells. Whereas ECE-1a and 1c were present at the plasma membrane, ECE-1b and ECE-1d were retained inside the cells. We found that both intracellular isoforms were concentrated in the endosomal system: ECE-1d in recycling endosomes, and ECE-1b in late endosomes/multivesicular bodies. Leucine-based motifs were involved in the intracellular retention of these isoforms, and the targeting of ECE-1b to the degradation pathway required an additional signal in the N terminus. The concentration of ECE-1 isoforms in the endosomal system suggested new functions for these enzymes. Potential novel functions include redistribution of other isoforms through direct interaction. We have showed that ECE-1 isoforms could heterodimerize, and that in such heterodimers the ECE-1b targeting signal was dominant. Interaction of a plasma membrane isoform with ECE-1b resulted in its intracellular localization and decreased its extracellular activity. These data demonstrated that the targeting signals specific for ECE-1b constitute a regulatory domain per se that could modulate the localization and the activity of other isoforms.

Acetylcysteine↗

Function of CBFbeta/Bro proteins.

Mammalian core binding factor beta (CBFbeta) and Drosophila Brother (Bro) and Big-brother (Bgb) proteins are transcription factors that dimerize with mammalian Runx and Drosophila Runt and Lozenge proteins and augment their DNA binding affinity and transcriptional potency. CBFbeta is essential for development and sustenance of definitive hematopoiesis during mouse embryogenesis. Bro and Bgb are required for Runt/Lozenge functions in Drosophila development. CBFbeta contributes to leukemogenesis since the CBFB gene is specifically and consistently mutated by a chromosome 16 inversion found in patients with acute myeloid leukemia subtype M4Eo. The ubiquitous expression pattern of the CBFB gene suggests that it may play important roles in many other organ systems.

Amino Acid Sequence↗

A selectable system for mutation detection in the Big Blue lacI transgenic mouse system: what happens to the mutational spectra over time.

Transgenic animals offer a powerful tool to study the mechanisms of spontaneous and induced mutagenesis in vivo. Herein we used a test version of a growth selectable assay to obtain spontaneous mutants in a lacI target transgene recovered from lacI transgenic B6C3F1 mice (Big Blue). This selection system may have certain advantages relative to the more established plaque screening system for mutation detection because: (1) the plating density of the phage is up to 60 times higher in the selectable assay, reducing the number of plates needed to be screened for a comparable amount of mutants; and (2) the mutant frequency obtained from the selectable assay is higher compared to the plaque assay, possibly due to a higher sensitivity for weaker mutants. However, the longer incubation time of the growth selectable assay might allow E. coli host derived mutants to appear. To address this issue, we investigated the sequence changes in the amino-terminal domain of the lacI gene of 405 mutants derived from the liver, spleen, brain, germ cells and skin of five untreated 6-week-old mice. The mutant colonies were isolated after 60, 84, 108 and 150 h of incubation under growth selectable conditions. Tissue-specific differences in the mutational pattern obtained after 60 and 84 h disappear after a longer time of incubation, possibly due to an increasing contribution of E. coli derived mutants. The evolving selectable systems offer the potential to increase screening efficiency, but the results suggest caution in interpreting data from this system because repair by E. coli of DNA lesions or mismatched heteroduplexes either originating in mouse in vivo or produced by ex vivo manipulation as well as de novo mutations in E. coli might contribute significantly to the observed mutational spectra at each timepoint.

Animals↗

Income as mediator of the effect of occupation on the risk of myocardial infarction: does the income measurement matter?

AIM: To investigate whether the effect of occupational grade on the risk of myocardial infarction (MI) is mediated by income with different aspects of income taken into account. METHODS: Data were used from three prospective population studies conducted in Copenhagen. A total of 16 665 employees, 43% women, aged 20-75 years, with an initial examination between 1974 and 1992 were followed up until 1999 for incident (hospital admission or fatal) MI. Register based information on job categories and income was used. RESULTS: During follow up, 855 subjects were diagnosed with a MI: 708 men and 147 women (in total 47% fatal). The hazards by household and individual income showed a graded effect with a hazard ratio (HR) of 1.43 (95% CI 1.12 to 1.83) for the lowest household income group compared with the highest, whereas equivalent income showed an inverse "J shape" effect with a HR of 1.55 (95% CI 1.25 to 1.82) for the third income group compared with the highest. HR for unskilled workers as compared with executive managers was reduced from 1.55 (95% CI 1.24 to 1.93) to 1.42 (95% CI 1.12 to 1.81) after adjustment for household income. CONCLUSIONS: Occupation and income are not mutually exclusive, but at least partly explained by or mediated through the other on the risk of MI. The mediating effect of income is independent of the choice of an income indicator. Income is not a big contributor to inequality in MI; probably because of the rather even income distribution in Denmark.

Adult↗

Sensitization to horse allergens without apparent exposure to horse. Report of two cases.

It has been well established that occupational exposure to horses increases the risk of allergic sensitization to horse hair, on the contrary very few data have been published in individuals without direct contact with horses. We describe two cases of severe respiratory symptoms triggered by a single direct exposure in two patients with only 4-5 previous short contacts with horses during the entire their life, since they live in urban area of Naples. Cutaneous and serological diagnostic tests showed significant sensitization to horse dander other than some common inhalant allergens. Our report suggests that, in already (but unaware) horse-sensitized patients, the last direct contact with a horse has triggered severe respiratory symptoms. Passive carriage of horse allergen by the clothes of individuals in contact with horses/horse environments or a cross-reactivity between horse allergen (a lipocalin) and the major allergens (also lipocalins) of several furred animals are likely to explain our findings. Some other considerations can be derived by the clinical observation. First, it is likely that allergic sensitization to horse allergens could be more frequent than expected also in subjects without direct exposure to horse because of living in urban areas. Secondly, individuals suspected to be highly atopic or those already sensitized to some pet dander should be tested by SPTs/radio-allergo-sorbent test before the introduction of a pet indoors or, in the case of a big animal as the horse, a direct horse contact/entering into environments containing a horse.

Adolescent↗

Worldwide prevalence of lentivirus infection in wild feline species: epidemiologic and phylogenetic aspects.

The natural occurrence of lentiviruses closely related to feline immunodeficiency virus (FIV) in nondomestic felid species is shown here to be worldwide. Cross-reactive antibodies to FIV were common in several free-ranging populations of large cats, including East African lions and cheetahs of the Serengeti ecosystem and in puma (also called cougar or mountain lion) populations throughout North America. Infectious puma lentivirus (PLV) was isolated from several Florida panthers, a severely endangered relict puma subspecies inhabiting the Big Cypress Swamp and Everglades ecosystems in southern Florida. Phylogenetic analysis of PLV genomic sequences from disparate geographic isolates revealed appreciable divergence from domestic cat FIV sequences as well as between PLV sequences found in different North American locales. The level of sequence divergence between PLV and FIV was greater than the level of divergence between human and certain simian immunodeficiency viruses, suggesting that the transmission of FIV between feline species is infrequent and parallels in time the emergence of HIV from simian ancestors.

Amino Acid Sequence↗

Role of endothelin in mechanisms of local blood pressure control.

A potent endothelium-derived vasoconstrictive peptide, endothelin (ET), is processed from a precursor, big endothelin, by an unknown converting enzyme. It stimulates calcium channels, phospholipase C and phospholipase A2 in smooth muscle cells through activation of the receptor specific for ET. The intracellular free calcium ion concentration is increased by ET, inducing vasoconstriction. The response to ET varies according to the type of blood vessel. Mesenteric small arteries and arterioles are sensitive to ET. It is likely that various factors, including adrenaline, angiotensin II and arginine vasopressin, induce synthesis and release of ET from the endothelium. A balance among ET, endothelium-derived relaxing factor (EDRF) and atrial natriuretic peptide (ANP) may control vascular tone and regulate peripheral blood flow.

Amino Acid Sequence↗

DNA adduct formation and oxidative stress in colon and liver of Big Blue rats after dietary exposure to diesel particles.

Exposure to diesel exhaust particles (DEP) via the gastrointestinal route may impose risk of cancer in the colon and liver. We investigated the effects of DEP given in the diet to Big Blue rats by quantifying a panel of markers of DNA damage and repair, mutation, oxidative damage to proteins and lipids, and antioxidative defence mechanisms in colon mucosa cells, liver tissue and the blood compartment. Seven groups of rats were fed a diet with 0, 0.2, 0.8, 2, 8, 20 or 80 mg DEP/kg feed for 21 days. DEP induced a significant increase in DNA strand breaks in colon and liver. There was no effect on oxidative DNA damage (8-oxodG) in colon or liver DNA or in the urine. However, the mRNA expression of OGG1, encoding an enzyme involved in repair of 8-oxodG, was increased by DEP in both liver and colon. DNA adduct levels measured by 32P-post-labelling were elevated in colon and liver, and the expression of ERCC1 gene was affected in liver, but not in colon. In addition to these effects, DEP exposure induced apoptosis in liver. There was no significant change in mutation frequency in colon or liver. The levels of oxidative protein modifications (oxidized arginine and proline residues) were increased in liver accompanied by enhanced vitamin C levels. In plasma, we found no significant effects on oxidative damage to proteins and lipids, antioxidant enzymes or vitamin C levels. Our data indicate that gastrointestinal exposure to DEP induces DNA adducts and oxidative stress resulting in DNA strand breaks, enhanced repair capacity of oxidative base damage, apoptosis and protein oxidation in colon mucosa cells and liver.

Animals↗

Cloning of full-length preproendothelin-2 cDNA and its expression in dog.

Endothelin-2 (ET2) is a member of the endothelin family of 21-amino acid peptides with vasoconstrictive activity. We report here the molecular cloning of the canine full-length cDNA of the precursor form of ET2, prepro-ET2 (PPET2), from intestinal tissue by means of reverse transcription-polymerase chain reaction (RT-PCR) in conjunction with 5'- and 3'-rapid amplification of cDNA ends (RACE). Aside from the poly (A) tail the cDNA was found to be 1195 bp and included an open reading frame of 534 bp encoding a PPET2 polypeptide of 178 residues, in which the regions corresponding to bioactive mature ET2 peptide, an intermediate form big-ET2, and endothelin-like peptide are found. The organ distributions of PPET2 mRNA and a splicing variant were analyzed by RT-PCR. PPET2 transcript was detected in duodenum, colon, stomach, lung, liver, uterus, ovary, testis and kidney, but not in spleen. A splicing variant was found in none of the organs. Thus, based on the cloned cDNA sequence, we established a quantitative assay for dog PPET2 mRNA level using a real-time PCR system. Quantitative analysis by this method in various organs of the dog demonstrated that the dominant gene expression occurs in the intestine, with higher expression in large intestine than in small intestine.

Animals↗

[Evaluation of different 16-row CT colonography protocols using a porcine model].

PURPOSE: To develop and to test an easily produced biological colon model with simulated polypoid lesions. Application of this phantom for the selection of an optimized scan protocol of 16-row CT colonography (CTC) for clinical use. METHODS AND MATERIAL: Six polypoid lesions (1 - 6 mm) were simulated with sutures on the inner face of a porcine colon segment (20 cm). After distending the colon segment with air, the phantom was placed in a water quench and CT scans were performed on a MDCT-scanner (Somatom Sensation 16, Siemens, Forchheim). At constant values for collimation (16x0.75 mm) and voltage (120 kV), 54 different combinations of mAs values (50, 75 and 100 mAs), pitch factors (1, 1.25 and 1.5) and slice thicknesses (0.75, 1.0, 1.5, 2.0, 3.0 and 5.0 mm) were tested systematically. The phantom was scanned in the longitudinal and transverse axis to simulate the different orientation of the colon in the abdomen. Axial slice images and virtual endoscopic views of all data sets were presented separately to 2 radiologists who independently determined number and size of detectable polyps. Dose exposure was measured with an Alderson phantom. RESULTS: The colon model offered a realistic imitation of a polyp-covered, human colon. The experimental set-up allowed a systematic evaluation of polyp detection related to lesion size, orientation of the colon and CTC parameters, with other influencing factors mostly excluded. Polyps were significantly better detected in the longitudinal than in the transverse orientation of the colon. For the detection of lesions of at least 3 mm, a low dose (50 mAs) 16-row CTC should be combined with a pitch of 1.5 and a maximum slice thickness of 3 mm. For the depiction of polyps smaller than 3 mm, slice thickness and pitch should amount to 1 mm and 1.0, respectively. Effective dose of this low dose protocol is 4.08 mSv. CONCLUSION: The porcine colon phantom represents a realistic and easily produced alternative to other colonography models. It allows a preselection of a CTC-protocol for subsequent clinical testings. If the high in vitro performance of the low-dose 16-row CTC-protocol is confirmed on a human collective, the use of 16-row technique would represent a big step for CTC toward a screening method.

Animals↗

[Anatomy, variations and malformations of the vessels of the head and neck region (author's transl)].

After a short review of developmental data concerning the head and neck arteries, the normal conditions, variations and malformations of single arteries are described. Special attention is paied to the skeletotopic and topographic relations of the arteries. In every bigger vessel its length, position, origin and branches are described, and in most cases are also demonstrated by diagrams and tables. As the aortic arch may also reach the neck region, its variation and malformations as well as the variations of its branches are mentioned. The variations and the malformations of the inominate artery, of the common carotid artery, as well as of the external and internal carotid arteries are described. The practical importance of these variations is stressed. Upon description of variations and malformations anatomical, surgical und roentgenological findings described in literature, as well as personal experiences are considered. Special attention has been paied to the variations of the circulus arteriosus Willisi, to the persistence of the presegmental arteries and to the possibility of formation of a by-pass in occlusion or aplasia of cerebral arteries. The position of the subclavian artery, its variations and the variations of its branches are described. After a short embryological introduction, the variations of the venous system and its anastomoses are described. At the end, the main variations of the big lymphatic vessels in the neck are mentioned.

Aorta, Thoracic↗

[A food cognition questionnaire].

BACKGROUND: Many authors evoke the role of cognition in the persistence of symptoms or in relapse. In pathology the cognitions produced by the patients are called dysfunctional or erroneous. The content of the cognitions are words or images issued from the treatment of information. In emotional disorders, the structure of thoughts named dysfunctional "schemata" involves a biased treatment of information and leads to erroneous cognitions. Several studies have attempted to elicit the most specific cognitions of different diseases. In this field, Hollon and Kendall found 36 cognitions specific to depression gathered in the automatic thoughts questionnaire (ATQ). In the same spirit, Beck et al. gathered 14 cognitions of anxiety and 12 depressed cognitions in the cognition check list (CCL). In the etiology and maintenance of eating disorders the cognitions take a large place. Around 1980 cognitive dysfunctioning was described and concerned food, interpersonal relationship and body shape. A few years later, some experimental studies explored these processes. The Stroop test, a categorization task, showed specific cognitive impairment in with patients eating disorders versus normal control subjects. It was then established that cognitive errors were based on food cognitions in restrictive patients, whereas they were based on body shape cognitions in bulimic patients. In several famous papers, Garner described typical cognitions of eating disorder patients and distinguished food-cognitions, eating-cognitions using case reports. As far as we know there is no clinical tool concerning such cognitions in France. That is the main motivation of the authors. AIM OF THE STUDY: The aim of this paper was to determine the characteristic cognitions of anorexic, anorexic-bulimic and bulimic patients and to compare them with those of normal control subjects. The goal of the study was to create a food cognition questionnaire. FIRST STEP METHODS: In the first step, food cognitions were collected among female eating disorder patients and normal female control subjects during systematic investigation. Ninety-two women were assessed and provided more than 3 000 food cognitions. Two independent psychologists identified the most frequent cognition per group and thus retained 115 food items. These items were randomly assigned. This provided the questionnaire. To illustrate the latter, here are the first five items: 1) Apricots are good for the health because they are rich in vitamins. 2) Pears are big fruit, difficult to digest. 3) Canned fruit is soaked with sugar. 4) Banana is a fruit which makes one put on weight. 5) White coloured food give the impression that it is not alive... The list of possible answers was: never, rarely, sometimes, often enough, often, always. SECOND STEP METHODS: In the second step, the food cognition questionnaire was proposed to 217 women including 131 eating disorder patients (53 anorexic, 50 anorexic bulimic, 28 bulimic) and 86 normal control subjects. The values of body mass index and the eating attitude test differed when we compared the two groups, and the mean age was close to 26 years in both groups. RESULTS: The statistic analysis highlighted six discriminative variables: two clinical criteria (weight and height) and four food-items given below: Q24: When I see food being fried, I feel the grease all over my body. Q76: When I start a cookie packet, I eat it up. Q102: When I feel anxious, I crave for food to fill my body. Q106: Eating pastry gives me heart-burn and makes me belch. The statistical model allowed us to differentiate eating disorder patients from normal control subjects. The content of the four food items is in agreement with experimental and clinical data. All these items included some aspects of the quality or quantity of food and also the negative consequences of food consumption on the body. CONCLUSION: To conclude, the model can help clinicians identify the patients and then initiate treatment. We also insist on the fact that this study is new and empirical, and should be extended by determining some food items for example, which would clarify the difference of behaviour between anorexics and bulimics.

Adult↗

[Ageing, genetics and the somatotropic axis].

Research on ageing made a big leap forward when genes regulating lifespan were discovered about a decade ago. First isolated by screening the genome of the nematode Caenorhabditis elegans, most of these genes belong to an essential signalling pathway that is highly conserved during animal evolution. Orthologous genes in vertebrate species are the families of genes coding for insulin, insulin-like growth factors (IGF) and related proteins. Intensively studied and well-known for their pivotal roles in proliferation, differentiation, survival and metabolism of most cells, we now discover their multiples functions with respect to the control of longevity and their ability to modulate the cell's responses to oxidative stress, a major cause of cellular and organismal ageing. The activity of IGF signalling in mammals depends on a complex interplay of endocrine signals that together constitute the somatotropic axis. Accordingly, several components of this hormone axis, like growth hormone or growth hormone releasing hormone receptors, regulate efficiently animal longevity, which has been elegantly demonstrated by studies performed in genetically modified mouse models. From this and other work, it becomes increasingly clear that the control of ageing is a question of hormonal regulations. We here present several of these models and discuss the respective contributions of insulin and IGF signalling to the regulation of lifespan. We review data on the Klotho gene that acts on lifespan via surprising and not yet fully understood molecular mechanisms, connecting this new, hormone-like substance to IGF and insulin signalling. We further report recent evidence showing that human lifespan might be controlled in similar ways. Finally, we shed some light on clinical GH treatment in humans, from an endocrinologist's point of view.

Aging↗

ETA receptor-mediated responses to endothelin-1 and big endothelin-1 in the rat kidney.

1. Renal clearance experiments were conducted in anaesthetized Sprague-Dawley rats to determine the effect of the ETA receptor antagonist, BQ-123, on the renal haemodynamic response to endothelin-1 (ET-1) and its precursor, big endothelin-1 (big ET-1) at doses that produce an equivalent degree of renal vasoconstriction. 2. Infusion of either big ET-1 at 100 pmol kg-1 min-1 or ET-1 at 12 pmol kg-1 min-1 for 60 min produced almost identical decreases in renal blood flow (RBF) and glomerular filtration rate (GFR). Big ET-1 produced an increase in mean arterial pressure (MAP) that was significantly larger than the increase produced by ET-1. 3. Co-infusion with BQ-123 (0.1 mg kg-1 min-1) prevented the rise in MAP produced by big ET-1 and completely blocked the renal response. Similarly, BQ-123 inhibited both the increase in MAP and the decrease in RPF and GFR produced by ET-1. 4. Big ET-1 but not ET-1, produced a significant increase in water and sodium excretion. BQ-123 had no effect on the diuretic and natriuretic response to big ET-1 consistent with possible ETB-mediated inhibition of tubular reabsorption. 5. We have previously shown that at higher doses of ET-1, BQ-123 was unable to inhibit the renal vasoconstrictor response despite blockade of the pressor response. Taken together, these results indicate that ET-1 activates primarily ETA receptors at moderately low doses to produce renal vasoconstriction while higher doses also involve non-ETA receptors. 6. Since ET-l has a similar binding affinity for both ETA and ETB receptors, we suggest that there maybe a third type of ET-1 receptor in the rat kidney with lower affinity for ET-1 that is coupled to a vasoconstrictor mechanism.

Amino Acid Sequence↗

Sex hormone-induced mammary carcinogenesis in female Noble rats: detection of differentially expressed genes.

Breast cancer is the most common cancer and the second most frequent cause of cancer death in women. Epidemiological data has recognized that an increased cumulative exposure to estrogen is the common tie linking most of the established risk factors for breast cancer. Sex hormone-induced mammary gland carcinogenesis of the Noble rat (using testosterone and 17beta-estradiol) resembles that of the human counterpart in its growth pattern as well as the histopathology of the tumors induced. This model may provide a paradigm for examination of genetic alterations and changes in gene expression between different histological groups and to make inferences about the role of known and putative oncogenes and tumor suppressor genes. We studied the gene expression profile during sex hormone-induced mammary carcinogenesis using a cDNA array technique; the results were further confirmed by RT-PCR, western blotting and immunohistochemical analyses. From the 10 differentially expressed genes identified, we have studied four highly overexpressed genes, two cell cycle/growth control regulators, the cyclins D1 and D2, a growth factor, IGF-2 and a cytokine TNF-alpha. Cyclins D1 and D2 were highly expressed in the nuclei of carcinoma cells but at low levels in the nuclei of the hyperplastic and normal mammary tissue. IGF-2 was found to expressed in the cytoplasm of the carcinoma cells but not in the stromal cells. Western blot showed expression of big IGF-2 consistent with the tumor derived truncated forms of pro-IGF-2. The matured circulating IGF-2 at 7.5 kDa identified in the serum was not expressed in any of the breast tissue samples. TNF-alpha expression was found not only in the macrophages but also in the mammary carcinoma cells. The result of the present study provides some information on the molecular basis of this sex hormone-induced mammary carcinogenesis and the role of these proteins in tumor progression.

Animals↗