RESUMO
Objective Whether the Ubi-p63E gene regulates spermatogenesis and tumorigenesis remains unclear. This study aimed to explore the regulatory effect of Ubi-p63E on germline stem cells (GSC) in the GSC niche of the Drosophila testis. Methods We used the UAS-Gal4 system for knockdown of the Ubi-p63E gene in the specific GSCs of the Drosophila testis and divided the male flies for this experiment into three groups: control (wild-type W1118 flies), nos>Ubi-p63E RNAi (knockdown of the Ubi-p63E gene in the early germ cells), and tj>Ubi-p63E RNAi (knockdown of the Ubi-p63E gene in the cystoblasts). We determined the fertility rate of the flies by fertility tests and examined the effect of Ubi-p63E on the Drosophila testis in the GSC niche by immunofluorescence staining. Results Fertility tests manifested a significantly lower rate of fertility in the nos>Ubi-p63E RNAi and tj>Ubi-p63E RNAi groups than in the control (0.00% and 4.12% vs 97.26%, P < 0.01). Morphologically abnormal testes were observed in the nos>Ubi-p63E RNAi and tj>Ubi-p63E RNAi groups, only 22.77% and 18.86% as long as the testes of the control flies. Immunofluorescence staining revealed no morphologically normal testes in the tj>Ubi-p63E RNAi group, but quite a few masses of abnormal cells, and mostly Vasa-positive cells. Conclusion The Ubi-p63E gene affects the self-renewal ability of GSCs in the GSC niche of the Drosophila testis as well as the differentiation of GSCs via cystoblasts, and consequently leads to the formation of germ cell tumors.
RESUMO
Objective Reports are rarely seen on the role of autophagy in missed abortion. This study aimed to explore the association of missed abortion with autophagy by detecting the expressions of the autophagy-related factors Beclin-1 and LC3 in the villus and decidua of normal pregnancy and missed abortion women so as to provide some theoretical evidence for the early prevention of missed abortion.Methods We included in the experimental group 30 missed abortion patients treated in the Department of Obstetrics and Gynecology, the Fourth Affiliated Hospital of Jiangsu University, from January to March 2017 and enrolled another 30 normal pregnancy women as controls. We determined the expression level of Beclin-1 mRNA in the villus and decidua by quantitative fluorescence PCR and those of Beclin-1 and LC3II/LC3I proteins by Western blot. We also positioned Beclin-1 in the villus and decidua tissues of the two groups of women by immunohistochemistry.Results Compared with the normal pregnancy women, the patients with missed abortion showed significantly up-regulated expression of Beclin-1 mRNA in the villus (1.00±0.28 vs 2.17±0.87, P<0.05) and decidua (1.75±0.41 vs 4.74±0.93, P<0.05) and those of Beclin-1 and LC3II/LC3I proteins (P<0.05). Immunohistochemistry revealed markedly increased expression of the Beclin-1 protein in the villus and decidua of the missed abortion patients (P<0.05).Conclusion Beclin-1 mRNA, Beclin-1 protein and LC3II/LC3I are all up-regulated in the villus and decidua of missed abortion patients, suggesting the involvement of autophagy in the occurrence of missed abortion, which may provide a new theoretical basis for studying the pathogenesis of missed abortion.