Upregulation of long noncoding RNA FERRE promoted growth and invasion of breast cancer through modulating miR-19a-5p/EZH2 axis.

OBJECTIVE: It has been demonstrated that long non-coding RNA (LncRNA) plays an important regulatory role in a series of diseases. The purpose of this study is to investigate the expression of long non-coding RNA (LncRNA) FERRE and its facilitating effects on proliferation and invasion of breast cancer by regulating oncogene EZH2 through sponging with miR-19a-5p. PATIENTS AND METHODS: qRT-PCR was performed to detect the expressions of FERRE and EZH2 in human breast cancer tissues and cells. CCK-8 assay was performed to evaluate the MCF-7 cells proliferation and transwell assay was performed to evaluate the MCF-7 cells migration. Correlation analysis between FERRE and miR-19a-5p was detected by statistical analysis. Bioinformatics prediction was made to detect the binding site of FERRE and miR-19a-5p and Luciferase activity was conducted to investigate the interaction between EZH2 and miR-19a-5p. Furthermore, we cloned the mice EZH2 3'-UTR into the Luciferase reporter vector and constructed miR-19a-5p binding mutants to validate the inhibited modulation of miR-19a-5p to the EZH2 expression. RESULTS: Results showed that expression of FERRE and EZH2 were upregulated in human breast cancer tissues and cells. qRT-PCR and CCK-8 assay showed that FERRE expression is associated with the proliferation of breast cancer cells, upregulated FERRE contributed to cell proliferation of MCF-7. Transwell assay showed that FERRE was associated with the migration ability of tumor cells, increased expression of FERRE promoted the migration and invasion of breast cancer cells. The bioinformatics prediction and Luciferase assay demonstrated that by sponging with miR-19a-5p, FERRE can serve as a molecular sponge to further regulate the expression of EZH2. CONCLUSIONS: We found that lncRNA-FERRE was upregulated in human breast cancer patients, which could accelerate tumor proliferation, migration and invasion as a molecular sponge by modulating the inhibitory effect of miR-19a-5p on oncogene EZH2.

Quantification of hemoglobin Constant Spring in heterozygote and homozygote by a capillary electrophoresis method.

INTRODUCTION: Capillary electrophoresis (CE) is a high-resolution method for detection of hemoglobin Constant Spring (Hb CS). METHODS: The levels of Hb CS quantified by CE were compared among three groups of samples including heterozygote and homozygote of Hb CS as well as Hb H-CS disease classified by DNA molecular diagnosis. The mean corpuscular volume (MCV) of red blood cells was also analyzed among these three groups. RESULTS: Mean ± SD of Hb CS level of the homozygote was not significantly different from that of the Hb H-CS disease (1.9 ± 1.8 vs. 2.8 ± 1.3, P = 0.13), but it was significantly higher than that of the heterozygote (1.9 ± 1.8 vs. 0.4 ± 0.2, P = 0.007). The MCV <70 fL was found in Hb H-CS disease only. CONCLUSION: CE is the preferable method for screening of heterozygote and homozygote of Hb CS. Moreover, in conjunction with a lower MCV (<70 fL), this approach provided a high resolution to identify Hb H-CS disease.

Effect of haematological alterations on thalassaemia investigation in HIV-1-infected Thai patients receiving antiretroviral therapy.

OBJECTIVES: To evaluate the effect of haematological alterations resulting from antiretroviral therapy (ART) on the diagnosis of thalassaemia carriers in HIV-1-infected Thai patients. METHODS: Complete blood cell counts, osmotic fragility (OF) test and haemoglobin (Hb)-A(2) values were measured in blood samples of 52 antiretroviral-treated and 14 untreated HIV-1-infected patients. Data were analysed according to thalassaemia type and ART. RESULTS: Sixteen patients carried at least one of the investigated thalassaemia types and most of them (87.5%) received ART. Their red cell indices [mean corpuscular Hb (MCH), mean corpuscular Hb concentration (MCHC) and red blood cell distribution width (RDW)], OF test and Hb-A(2) values were observed within the critical criteria of each thalassaemia type. Normocytic red cells were observed in alpha-thalassaemia and Hb-E trait. Among HIV-1-infected patients who are non-thalassaemia carriers, higher values of Hb-A(2), MCH, macrocytosis and lower red cell counts were observed in the treated group. Values of RDW, MCHC and OF test for treated and untreated groups were in the normal range. Five treated patients had Hb-A(2) values within the critical criteria of beta-thalassaemia carriers but beta-thalassaemia gene mutations were not observed by polymerase chain reaction analysis. CONCLUSIONS: ART can alter many haematological figures. Therefore, diagnosis of thalassaemia should be evaluated carefully in combination with those parameters.

Characterization of the main placental cytokine profiles from HIV-1-infected pregnant women treated with anti-retroviral drugs in France.

Cytokines are involved in regulating HIV-1 infection. They are also placental environment major components. We assessed the potential impact of HIV-1 infection and/or anti-retroviral drugs on the placental cytokine profiles that may be involved in controlling HIV-1 placental dissemination. Placental explants were obtained after elective caesarean section from anti-retroviral-treated HIV-1-infected pregnant women and from HIV-1 non-infected pregnant women. The main placental cytokines were assessed for protein secretion in the supernatants of 24-h placental culture explants and/or in uncultured placental explants for mRNA expression levels. The cytokine profiles were different between the HIV-1-infected and the non-infected groups. Higher medians of leukaemia inhibiting factor (LIF), tumour necrosis factor (TNF)-alpha and interleukin (IL)-8 secretion were found in the 24-h culture supernatant of term placenta from HIV-1-infected women. High median levels of IL-16 and regulated upon activation normal T cell expressed and secreted (RANTES) levels were found in both groups. The mRNA expression medians were lower for TNF-alpha and IL-8 and higher for stromal cell-derived factor-1 (SDF-1) in uncultured placental explants from HIV-1-infected women. In the HIV-1-infected group, but not in the non-infected group, the secretion levels of TNF-alpha and IL-8, as well as their mRNA expression levels, were highly positively correlated; furthermore, their secretion levels were correlated positively with LIF and IL-10 secretion levels. We found no correlation between the cytokine levels and the immunovirological status of the HIV-1-infected mothers or the type or duration of treatment. These results highlight the potential impact of HIV-1 and of the anti-retroviral treatments on the placental cytokines pattern, independently of their anti-viral activity.

Down modulation of TNF-alpha mRNA placental expression by AZT used for the prevention of HIV-1 mother-to-child transmission.

Mechanisms of HIV-1 in utero mother-to-child transmission (MTCT) protection provided by AZT are not completely understood. The placental cytokine network is involved in the control of HIV-1 in utero transmission but the effect of AZT on this network is unknown. To evaluate the effects of AZT on placental cytokine expression, the chorionic villi from HIV-1 uninfected women term placentae were cultured with 0, 100, and 2,000 ng/ml AZT. Tissue fragments were harvested at days 1, 4, and 7 to determine the level of cytokine mRNA by real-time RT-PCR. The viability and morphology of the placental histocultures were monitored by the expression of beta-human chorionic gonadotropin (beta-hCG) gene, lipopolysaccharide (LPS) activation, and microscopic examination. AZT at 2,000 ng/ml significantly down-regulated TNF-alpha mRNA expression at day 1 and day 4, but had no effect on beta-hCG, stromal cell-derived factor 1 (SDF-1), and IL-10 gene expression. AZT did not induce any deleterious impact on placental tissue structure. Furthermore, activation of chorionic villi by LPS for 24 h up-regulated IL-10 and TNF-alpha mRNA expression. Down-regulation of TNF-alpha mRNA could represent a mechanism through which AZT can decrease the risk of HIV-1 MTCT, in addition to its direct effect on HIV-1 replication.

Evaluation of the placental environment with a new in vitro model of histocultures of early and term placentae: determination of cytokine and chemokine expression profiles.

We aimed to set up and validate a new in vitro model of placental histocultures, for the evaluation of cytokine and chemokine profiles of the placental environment, over a long culture period. Micro-explant cultures from 6 early and 6 term placentae were set up on collagen sponge gel supports at a liquid/air interface. At various times during culture, we analyzed tissue morphology and cell death by microscopy and quantified beta-hCG production and mRNA levels for beta-hCG and insulin-like 4 (INSL4). Levels of IL-6, LIF, TNF alpha, IL-10, IFN-gamma, IL-16 and RANTES in the medium were measured by ELISA on days 1, 4 and 7 of culture. SDF-1 mRNA expression was determined by real-time PCR at the same time points. Histocultures from early and term placentae remained viable until day 10. High levels of IL-6 and LIF production, low levels of TNF alpha, IL-10 and IFN-gamma production and significant SDF-1 expression were observed. These data indicate that placental histoculture is a suitable and reliable in vitro model for studying the placental environment.