The role of transmembrane protein 27 (TMEM27) in islet physiology and its potential use as a beta cell mass biomarker.

AIMS/HYPOTHESIS: Transmembrane protein 27 (TMEM27) is a membrane protein cleaved and shed by pancreatic beta cells that has been proposed as a beta cell mass biomarker. Despite reports of its possible role in insulin exocytosis and cell proliferation, its function in beta cells remains controversial. We aimed to characterise the function of TMEM27 in islets and its potential use as a beta cell mass biomarker. METHODS: To determine TMEM27 function, we studied TMEM27 gene expression and localisation in human healthy and diabetic islets, the correlation of its expression with cell cycle and insulin secretion genes in human islets, its expression in tungstate-treated rats, and the effects of its overproduction on insulin secretion and proliferation in a beta cell line and islets. To elucidate its utility as a beta cell mass biomarker, we studied TMEM27 cleavage in a beta cell line, islets and primary proximal tubular cells. RESULTS: TMEM27 mRNA levels in islets are lower in diabetic donors than in controls. Its gene expression correlates with that of insulin and SNAPIN in human islets. TMEM27 expression is downregulated in islets of tungstate-treated rats, which exhibit decreased insulin secretion and increased proliferation. TMEM27 overproduction in a beta cell line and islets significantly enhanced glucose-induced insulin secretion, with modest or no effects on proliferation. Finally, TMEM27 is cleaved and shed by renal proximal tubular cells and pancreatic islets. CONCLUSIONS/INTERPRETATION: Our data support a role for TMEM27 in glucose-induced insulin secretion but not in cell proliferation. The finding that its cleavage is not specific to beta cells challenges the current support for its use as a potential beta cell mass biomarker.

Effects of an androgenic derivative on pre-established mammary tumours chemically induced in the rat.

The effects were studied of an androgenic derivative--danazol--administered at doses of 10-12 mg kg-1 day-1 during 97 days to rats with dimethylbenz[a] anthracene-induced mammary tumours. Our main observations were as follows. (a) Danazol did not influence ovarian function at the end of the assay. (b) The treatment with danazol reduced the incidence (P less than 0.05), number of tumours (P less than 0.05) and volume of malignant mammary tumours; on the other hand, the values of these parameters for benign tumours and those of doubtful expression were similar in both experimental groups. (c) Such differential action of Danazol seems to be due to the different incidence and/or content of receptors of both types of tumours. (d) The latter results lead to a hypothesis for the mechanism of action of danazol based on its behaviour at different levels.

Effects of an androgenic derivative on the development of chemically-induced mammary carcinogenesis in the rat.

The effects of an androgenic derivative--danazol--on the development of dimethylbenz(a)anthracene (DMBA)-rat mammary carcinogenesis were studied. Animals in the treated group received danazol (10-12 mg/kg/day) during 169 days, starting 5 days after DMBA induction. As compared with tumours in control animals, those treated with danazol appear later and are significantly smaller. Moreover, this treatment reduced the incidence of animals affected by mammary cancer and the average number of malignant tumours in each animal. In the same way, the incidence of animals with missing nodules was significantly higher in the group treated with danazol. It is concluded that danazol has an inhibitory effect on experimental mammary tumours. This effect seems to be greater the earlier the treatment is started and the longer time it is applied.

Experimental diets for the study of lipid influence on the induced mammary carcinoma in rats: II--Suitability of the diets.

We have previously reported one method for obtention of experimental diets for the study of the effects of dietary lipids on the rat breast carcinoma. The purpose of this second part of the study was to develop a quality control system for demonstrating the suitability of these diets. This system is essentially based on the animals' growth control, their period clinical examination as well as the anatomopathological postmortem study of the animals submitted to such diets. Two groups of weaning rats, control (C) and hyperlipidic (HL), were submitted to a low-fat diet (N3) or a high-fat polyunsaturated--corn oil--diet (HL20) respectively. At 53 days of age all animals were induced with 5mg of dimethylbenz (a) anthracene. Experiments were ended when animals reached a mean age of 214 days. The results show: 1) a normal ponderal evolution of the animals in the two experimental groups with respect to two series of the same strain fed with a standard diet, and 2) the homogeneity of growth determined by the coefficient of variance study. On the other hand, neither the weekly clinical examination nor the anatomopathological post-mortem studies revealed any pathology that could be specifically attributed to nutritional imbalance. These results confirm the suitability of both diets for rat growth. Their use in the study of the effects of dietary lipids on the mammary carcinoma would satisfy the initial aim of guaranteeing the specificity of the results.

Dietary polyunsaturated n-6 lipids effects on the growth and fatty acid composition of rat mammary tumors.

The aim of this study was to analyze the effects of a polyunsaturated n-6 high-fat diet on rat DMBA-induced breast cancer at different stages of the carcinogenesis and to investigate if changes in the tumor fatty acid composition are one of the mechanisms by which dietary lipids could exert their effects. 14 fatty acids were evaluated in 6 lipid fractions. The results firstly showed that this high-fat diet stimulated the malignant mammary tumor growth, mainly all in the promotion group. The tumor lipid analysis indicated: 1) that each lipid fraction presented distinct major fatty acids (>5%) which were not the most abundant in the diet, except in the case of the triacylglicerides, suggesting the different resistance to dietary fatty acid modification of the tumor lipid fractions; 2) a higher arachidonic acid content in the fractions with less linoleic acid, above all in phospholipids, particularly in the phosphatidylethanolamine, indicating a different efficiency of conversion; 3) the three most abundant fatty acids in the dietary lipid (18:2n-6, 18:1n-9 and 16:0) were those which essentially displayed the differences between groups; thus, the high-fat diet changed the tumor lipid profile, increasing the 18:2n-6 relative content and decreasing that of the 18:1n-9; differences were significant in phosphatidylcholine, free fatty acids and triacylglycerides. Any change was obtained in the phosphatidylinositol. The greatest number of differences was found in the promotion group. Taken as a whole, our results suggest the different roles of lipid fractions in breast cancer cells and an association between cancer malignancy and the content of linoleic and oleic acids.