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| Author(s): |
Isabella Barbutti
Total Authors: 1
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| Document type: | Doctoral Thesis |
| Press: | Campinas, SP. |
| Institution: | Universidade Estadual de Campinas (UNICAMP). Instituto de Biologia |
| Defense date: | 2020-10-14 |
| Examining board members: |
Hernandes Faustino de Carvalho;
Alexandre Bruni Cardoso;
Luciana Bolsoni Lourenço;
Murilo Vieira Geraldo;
Vanessa Morais Freitas
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| Advisor: | Hernandes Faustino de Carvalho |
| Abstract | |
HBA and HBB genes cluster in different chromosomes. They are commonly expressed in cells from the erythroid compartment in a stoichiometric manner. Thus, their expression in this compartment is tightly regulated by a series of widely known factors. Nevertheless, there are several papers reporting haemoglobin expression in other tissues, some of them pointing to the gene regulation behind the expression and/or haemoglobin functionality in these foreign tissues. Interestingly, previous studies from our laboratory have found haemoglobin expression in rodent prostate cells: in castrated rats and in mice fed with normolipidic diets based on pork fat (lard). Additionally, published articles have described activity of the human HBG promoter in mice p63+ prostate cells and in human prostate cell lines. Thus, we intended to study the conditions in which haemoglobins are expressed in the prostate, as well as whether the gene regulation mechanisms are similar to that found in erythroid cells. Because haemoglobin expression in the prostate seems correlated with an undifferentiated state of the epithelial cells, we have also studied a possible link between haemoglobin expression and tumour progression. Therefore, we have characterized haemoglobin expression and subcelular localization in normal and tumour human epithelial prostate cell lines cultured bi- and three-dimensionally, and treated with androgens. We have also studied the effect of different concentrations of androgens in mice prostate, using castration and/or hormonal readministration, as well as the effects of aging, on haemoglobin expression. We concluded that normal epithelial prostate cell lines have higher haemoglobin expression when compared to tumour cells, and that dihydrotestosterone (DHT) signalling does not alter haemoglobin expression in vitro or in vivo. RWPE1 cell line expresses HBA2, HBA1 (alpha cluster) and HBE (beta cluster), and expression of HBA increases when cells are tridimensionally cultured. Moreover, HBA subcelular localization changes in some of the spheroid’s cells. Embryonic haemoglobins are the only ones from the ? cluster that are expressed, both in vivo (Hbb-by in aged mice prostate) and in vitro (HBE in RWPE1 cell line). Haemoglobin expression in prostate cells is not regulated in the same manner as in erythrocytes. Changes in haemoglobin expression found in the RWPE1 cell line also cannot be explained by DHT stimuli or p63 expression, which are specific features of prostate cells, nor by other mechanisms, such as extracellular matrix signalling or hypoxia, even though HBA is found overexpressed in the three-dimensional culture. (AU) | |
| FAPESP's process: | 16/06543-4 - Globin genes expression patterns and regulation mechanisms in the prostate gland |
| Grantee: | Isabella Barbutti Gonçalves |
| Support Opportunities: | Scholarships in Brazil - Doctorate |