Signal transducer and activator of transcription 3 (Stat3) is implicated in the pathogenesis of many malignancies and essential for IL-6–dependent survival and growth of multiple myeloma cells. Here, ...we demonstrate that the gene encoding oncogenic microRNA-21 (miR-21) is controlled by an upstream enhancer containing 2 Stat3 binding sites strictly conserved since the first observed evolutionary appearance of miR-21 and Stat3. MiR-21 induction by IL-6 was strictly Stat3 dependent. Ectopically raising miR-21 expression in myeloma cells in the absence of IL-6 significantly reduced their apoptosis levels. These data provide strong evidence that miR-21 induction contributes to the oncogenic potential of Stat3.
Background/ Aims: This study was performed to reveal signaling pathways exploited by pigment epithelium-derived factor (PEDF) derived from retinal (glial) Müller cells to protect retinal ganglion ...cells (RGCs) from cell death. Methods: The survival of RGCs was determined in the presence of conditioned culture media (MCM) from or in co-cultures with Müller cells. The significance of PEDF-induced STAT3 activation was evaluated in viability assays and using Western blotting analyses and siRNA-transfected cells. Results: Secreted mediators of Müller cells increased survival of RGCs under normoxia or hypoxia to a similar degree as of PEDF- or IL-6-exposed cells. PEDF and MCM induced an increased STAT3 activation in RGCs and R28 cells, and neutralization of PEDF in MCM attenuated STAT3 activation. Inhibition of STAT3 reduced PEDF-promoted survival of RGCs. Similar to IL-6, PEDF induced STAT3 activation, acting in a dose-dependent manner via the PEDF receptor (PEDF-R) encoded by the PNPLA2 gene. Ablation of PEDF-R attenuated MCM-induced STAT3 activation and compromised the viability of PEDF-exposed R28 cells. Conclusions: Müller cells are an important source of PEDF, which promotes RGC survival through STAT3 activation and, at least in part, via PEDF-R. Enhancing the secretory function of Müller cells may be useful to promote RGC survival in retinal neurodegenerative diseases.
Müller glial cells carry out different tasks to warrant normal retinal functions. The aim of this study was to investigate if Müller cells also support retinal ganglion cells (RGC).
RGC were cultured ...for 24 hours in the presence or absence of Müller glial cells under normoxic (20% O
, 5% CO
) or hypoxic (0.2% O
, 5% CO
, 94.8% N
) culture conditions. The number of vital RGC and the length of the newly developed neurites were evaluated.
Under normoxic conditions, RGC vitality was significantly higher (p < 0.01) when cultured with Müller cells (62.85 ± 2.06%) than without (47.29 ± 2.83%). Under hypoxia, RGC vitality was significantly higher (p < 0.01) in co-cultures (41.07 ± 2.28%) than in homotypic RGC cultures (28.49 ± 2.16%). The maximum length of the newly developed neurites was found in the normoxic co-culture (90.7 ± 7.4 µm), but showed only a minor difference (p = 0.04) when compared to the normoxic homotypic RGC culture.
Müller glial cells support RGC under normoxic and hypoxic culture conditions. Length of newly developed neurites and number of surviving RGC are both parameters to evaluate cell vitality.
SRC (steroid receptor co-activator)-1 has been reported to interact with and to be an essential co-activator for several members of the STAT (signal transducer and activator of transcription) family, ...including STAT3, the major signal transducer of IL (interleukin)-6. We addressed the question of whether SRC-1 is crucial for IL-6- and STAT3-mediated physiological responses such as myeloma cell survival and acute-phase protein induction. In fact, silencing of SRC-1 by RNA interference rapidly induced apoptosis in IL-6-dependent INA-6 human myeloma cells, comparable with what was observed upon silencing of STAT3. Using chromatin immunoprecipitation at STAT3 target regions of various genes, however, we observed constitutive binding of SRC-1 that decreased when INA-6 cells were treated with IL-6. The same held true for STAT3 target genes analysed in HepG2 human hepatocellular carcinoma cells. SRC-1-knockdown studies demonstrated that STAT3-controlled promoters require neither SRC-1 nor the other p160 family members SRC-2 or SRC-3 in HepG2 cells. Furthermore, microarray expression profiling demonstrated that the responsiveness of IL-6 target genes is not affected by SRC-1 silencing. In contrast, co-activators of the CBP CREB (cAMP-response element-binding protein)-binding protein/p300 family proved functionally important for the transactivation potential of STAT3 and bound inducibly to STAT3 target regions. This recruitment did not depend on the presence of SRC-1. Altogether, this suggests that functional impairment of STAT3 is not involved in the induction of myeloma cell apoptosis by SRC-1 silencing. We therefore conclude that STAT3 transactivates its target genes by the recruitment of CBP/p300 co-activators and that this process generally does not require the contribution of SRC-1.
Signal transducer and activator of transcription 3 (Stat3) is implicated in the pathogenesis of many malignancies and essential for IL-6–dependent survival and growth of multiple myeloma cells. Here, ...we demonstrate that the gene encoding oncogenic microRNA-21 (miR-21) is controlled by an upstream enhancer containing 2 Stat3 binding sites strictly conserved since the first observed evolutionary appearance of miR-21 and Stat3. MiR-21 induction by IL-6 was strictly Stat3 dependent. Ectopically raising miR-21 expression in myeloma cells in the absence of IL-6 significantly reduced their apoptosis levels. These data provide strong evidence that miR-21 induction contributes to the oncogenic potential of Stat3.
In the southeast of Bosnia and Herzegovina, the Berkovići earthquake sequence started with the mainshock on 22 April 2022 21:07 UTC at focal depth 22 km with magnitude ML = 6.0 (Mw = 5.7). Our ...preliminary estimation of the mainshock's maximum intensity is VII EMS for Berkovići where 29% of buildings were damaged. We analysed the first nine months of this sequence, 22 April 2022–22 January 2023. The earthquakes were located using a guided grid-search algorithm with source-specific station corrections as a mean of solutions for 54 combinations of velocity models and program control parameters. The analysis of aleatory variation and epistemic uncertainty showed that they are very dependent on the station coverage, especially for focal depth. The event catalogue consists of 7217 earthquakes and can be considered complete for ML ≥ 1.3. Focal depths (15–30 km) are considerably larger than average for the Dinarides, but consistent within the zone of mid-crustal events where the earthquakes occurred. Focal mechanisms were determined with the first-motion polarity method for eight earthquakes: five of them, including the mainshock, were due to reverse faulting on faults striking in the Dinaric direction, with the preferred main fault gently dipping to the northeast. However, three events were due to normal faulting, unexpected for this area. We constructed a regional seismotectonic cross-section to delineate a potential seismogenic source of the mainshock, and it suggests that the mainshock occurred on the NE-dipping blind ramp of the basal thrust of the Dalmatian tectonic unit. Moreover, another NE-dipping and blind ramp of this basal thrust could be responsible for the Ston–Slano 1996 earthquake, located to the SW of the Berkovići mainshock hypocentre at the horizontal distance of c. 35 km.
•The Berkovići earthquake sequence started with the mainshock on 22 Apr 2022 (21:07, ML = 6.0) at the focal depth of 22 km.•The earthquake catalogue (22 Apr 2022–22 Jan 2023) contains 7217 events with a completeness magnitude of 1.3.•Focal depths (15–30 km) are considerably larger than average for the Dinarides.•Focal mechanisms for three (of eight) earthquakes were due to normal faulting that is unexpected in this area.•Seismotectonic analysis hints the mainshock occurred on the blind ramp of the basal thrust of the Dalmatian tectonic unit.
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