Novel mechanism of miRNA-365-regulated trophoblast apoptosis in recurrent miscarriage.

Clinical pregnancies increasingly end in recurrent miscarriage (RM) during the first trimester, with genetic factors shouldering the main responsibility. MicroRNAs (miRNAs) regulate gene expression in a wide array of important biological processes. We examined the potential role of dysregulated miRNAs in RM pathogenesis and trophoblast development as an approach to elucidate the molecular mechanism behind RM. miRNA profiles from clinical specimens of RM and induced abortion (IA) were compared, and several miRNAs were found to be aberrantly expressed in RM samples. Among the miRNAs, miR-365 was significantly differentially expressed in RM decidual tissues. Furthermore, our results demonstrate that miR-365 functions as an upstream regulator of MDM2/p53 expression, cell cycle progression and apoptosis in trophoblasts. Bioinformatic prediction and experimental validation assays identified SGK1 as a direct target of miR-365; consistently, its protein levels were low in decidual tissues. Additionally, functional studies revealed that SGK1 silencing elicits cell cycle arrest and apoptosis in trophoblasts and that SGK1 overexpression attenuates the effects of miR-365 on apoptosis and MDM2/p53 expression. Collectively, our data provide evidence that the up-regulation of miR-365 may contribute to RM by decreasing SGK1 expression, which suggests its potential utility as a prognostic biomarker and therapeutic target for RM.

The role of globular heads of the C1q receptor in HPV 16 E2-induced human cervical squamous carcinoma cell apoptosis is associated with p38 MAPK/JNK activation.

BACKGROUND: Human papillomavirus type 16 (HPV 16) E2 protein is a multifunctional DNA-binding protein. HPV 16 E2 regulates many biological responses, including DNA replication, gene expression, and apoptosis. The purpose of this study was to investigate the relationship among the receptor for globular heads of the human C1q (gC1qR) gene expression, HPV 16 E2 transfection and apoptosis regulation in human cervical squamous carcinoma cells (C33a and SiHa). METHODS: gC1qR expression was examined in C33a and SiHa cells using real-time PCR and Western blot analysis. Apoptosis of C33a and SiHa cells was assessed by flow cytometry. C33a and SiHa cell viability, migration and proliferation were detected using the water-soluble tetrazolium salt (WST-1) assay, a transwell assay and 3H-thymidine incorporation into DNA (3H-TdR), respectively. RESULTS: C33a and SiHa cells that were transfected with a vector encoding HPV 16 E2 displayed significantly increased gC1qR gene expression and p38 mitogen-activated protein kinase (p38 MAPK)/c-jun N-terminal kinase (JNK) activation as well as up-regulation of cellular apoptosis, which was abrogated by the addition of gC1qR small interfering RNA (siRNA). Furthermore, the changes in C33a and SiHa cell viability, migration and proliferation that were observed upon HPV 16 E2 transfection were abrogated by SB203580 (a p38 MAPK inhibitor) or SP600125 (a JNK inhibitor) treatment. CONCLUSION: These data support a mechanism whereby HPV 16 E2 induces apoptosis by silencing the gC1qR gene or inhibiting p38 MAPK/JNK signalling in cervical squamous cell carcinoma.

[Changes of regulatory T cells and T helper cells in peripheral blood and their roles in the severity evaluation in children with asthma].

OBJECTIVE: To investigate the changes of CD4+CD25+Foxp3+ regulatory T cells (Treg) and T helper cells (Th1/Th2) in peripheral blood and their roles in the severity evaluation in children with asthma. METHODS: One hundred and fifty children with asthma were classified into acute attack (94 cases) and remission (56 cases) groups according to their clinical features, and the acute attack children were subdivided into mild asthma (54 cases) and severe asthma (40 cases) groups. Fifty healthy children were enrolled as a control group. The levels of CD4+CD25+Foxp3+ Treg, CD4+IFN-γ+ Th1 and CD4+IL-4+ Th2 in peripheral blood were measured by flow cytometer. RESULTS: The mean levels of CD4+CD25+Foxp3+ Treg and the ratio of Th1/Th2 in asthmatic children were lower than those in the control group (P<0.01). The Treg levels and the ratio of Th1/Th2 in the acute attack group were lower than those in the remission group and in the control group (P<0.01). The Treg levels in the severe asthma group were lower than those in the mild asthma group (P<0.01). There was a remarkably negative correlation between Treg levels and the asthma severity (r=-0.737, P<0.01), and the Th1/Th2 ratio was also negatively correlated with the asthma severity (r=-0.615, P<0.01). The Treg levels were positively correlated with the Th1/Th2 ratio (r=0.856, P<0.01). CONCLUSIONS: The Treg levels decrease remarkably and Th subsets imbalance occurs in children with asthma. This suggests that Treg and Th immunity play important roles in the pathogenesis of asthma. The Treg levels and the ratio of Th1/Th2 in peripheral blood may be useful in the evaluation of severity in children with asthma.

Regulatory T cells and Th1/Th2 in peripheral blood and their roles in asthmatic children.

OBJECTIVE: To determine the changes in CD4(+)CD25(+)Foxp3(+) regulatory T cells (Treg) and the Th1/Th2 ratio in peripheral blood of children with asthma, in order to investigate its association with asthma. METHODS: A total of 150 children with asthma were allocated into the acute phase group (n=94) and remission phase group (n=56) based on clinical manifestations. The acute phase group was subdivided into the mild group (n=54) and severe group (n=40). Flow cytometry was applied to determine CD4(+)CD25(+)Foxp3(+) Treg, CD4(+)IFN-γ(+) Th1 and CD4(+)IL-4(+) Th2 levels in peripheral blood of different groups, and the results were compared with normal children (control group, n=50). RESULTS: The Treg level was significantly lower in the asthma group than the control group (P<0.01): the Treg level was significantly lower in the acute phase group than the remission phase group and control group (P<0.01) and also significantly lower in the severe group than the mild group (P<0.01). The Th1/Th2 ratio was significantly lower in the asthma group than the control group (P<0.01) and also significantly lower in the acute phase group than the remission phase group and control group (P<0.01). The Treg level in peripheral blood of asthmatic children was negatively correlated with the severity of asthma (r=-0.737, P<0.01) and the Th1/Th2 ratio was also negatively correlated with the severity of asthma (r=-0.615, P<0.01), but the Treg level was positively correlated with the Th1/Th2 ratio (r=0.856, P<0.01). CONCLUSIONS: The significantly decreased level of Treg in peripheral blood and Th subset imbalance in asthmatic children suggest the important roles of Treg and Th immunity in pathogenesis of asthma. The Treg level and Th1/Th2 ratio in peripheral blood can be used to evaluate the severity asthma.