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PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
Before the experiments, the efficiency of the reaction for each pair of primers was checked.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
The cDNA was serially diluted 5-5000, qPCR was performed, and the efficiency of the reaction was calculated in MS Excel via the equation E = 10.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
E = 2 specifies 100% efficiency.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
Only starters for which the calculated reaction efficiency was greater than 95% were used in the experiments.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
Doxorubicin is a chemotherapeutic agent that is actively removed from cells by the ABCB1 efflux pump.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
The cells were treated with a 30 nM solution of doxorubicin (sc-200923, Santa Cruz) (dose selected on the basis of a previous publication or water (solvent) and then seeded at a density of 1 × 10 cells in 5 ml of medium.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
The mixture was incubated for 48 h at 37 °C, after which ATP and P21 protein levels were measured as described above.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
The experiments were performed three times.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
The ABCB1 efflux pump actively removes foreign substances from the inside of the cells.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
One such substance is rhodamine 123, which possesses fluorescent properties.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
The cells were seeded at a density of 1 × 10 cells in 5 ml of medium and incubated overnight.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
Next, the cells were treated for 4 h with a 1 µM solution of rhodamine 123 (83702, Sigma‒Aldrich), washed with PBS, and observed via a fluorescence microscope (Nikon Eclipse Ti-S, Japan).
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
The excitation and emission wavelengths were 485 and 539 nm, respectively.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
The experiments were performed three times.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
PHLDA1-silenced (shP) and control (shC) cells were grown in 150 cm culture flasks.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
The cells were harvested via TrypLeTM reagent (12604021, Gibco) incubation for 1–2 min at 37 °C.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
Afterwards, the enzyme was inactivated with 10% FBS in EMEM (with 1% NEAA, 1 mM sodium pyruvate) without the antibiotics gentamycin and puromycin, and the cells were subsequently centrifuged (200 × g, 5 min).
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
The pellets were resuspended in PBS.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
The pools of all resuspended cells were mechanically dispersed to dissociate clumps (as described in) and diluted in PBS for cell counting in a Bürker counting chamber.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
The excess cells were then rewashed with PBS (200 × g, 5 min).
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
After the removal of the PBS, fresh PBS was added, and the cells were recounted to a specific number for an experiment (with the appropriate excess).
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
Confirmation of PHLDA1 levels in modified cells prepared for injection into mice was performed via western blotting.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
ShP and shC IMR-32 cells (1 × 10 per well of a 6-well plate) were seeded and cultured for 48 h. Afterwards, the cells were harvested and lysed in RIPA buffer supplemented with protease and phosphatase inhibitors.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
After PHLDA1 detection, the membrane was stripped, and then α-tubulin was detected (Supplementary Fig. S14, S15 online).
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
To generate xenograft tumors, athymic nude mice (Rj: ATHYM-Foxn1nu/nu, Janvier Lab, France) were used on the basis of procedures approved by the 2nd Local Animal Ethical Committee in Kraków, Poland (Approval no. 153/2022).
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
All the experiments were performed in accordance with the relevant guidelines and regulations.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
The authors complied with the ARRIVE guidelines.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
The mice were housed under pathogen-free conditions in a temperature-controlled (approximately 22 °C) animal facility with a 14/10 hour light/dark cycle (in ventilated cages, with 55 ± 10% humidity) and were fed ad libitum.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
The mice were allocated to cages (4 animals per cage) by the animal care staff, but no other blinding/randomization was applied by the researchers.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
The animals were housed in the same rack and at the same height for a given experiment.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
No criteria for the exclusion or inclusion of animals were set a priori, and no animals were excluded during the described experiments.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
Each of the 2 separate experiments described here involved 16 mice in 2 groups, which were named after the treatment received.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
Each experiment included a control group.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
The mice (8 females per group, approximately 6 weeks old) were subcutaneously (s.c.)
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
injected without anesthesia in the right flank with 4 × 10 shP or shC IMR-32 cells in Matrigel Matrix HC (354262, Corning; the Matrigel was diluted 1:1 with cells in PBS, and the injection volume was 200 µl).
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
The s.c.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
tumors were monitored at the inoculation site via caliper measurements to ensure that the tumors did not exceed a volume of 1500 mm.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
The weight was monitored to avoid weight loss of more than 20%.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
At the endpoint, the animals were euthanized with carbon dioxide (CO2), and the tumors were collected, weighed, and processed immediately.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
The collected tumor samples were incubated for 24 h in 10% formalin (114321730, Chempur), dehydrated in increasing concentrations of ethanol and xylene, and then embedded in paraffin.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
The paraffin blocks were stored at 4 °C.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
The paraffin blocks were briefly frozen at -20 °C before being cut on a microtome.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
To examine the histology of the tumors, formalin-fixed, paraffin-embedded (FFPE) 4 μm tumor slices were stained with hematoxylin and eosin.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
To examine the collagen net within the tumors, FFPE slices were stained with a Masson trichrome kit (3459.1, Carl Roth GmbH + Co. KG) and a van Gieson Trichrome kit (8275.1, Carl Roth GmbH + Co. KG) according to the manufacturer’s protocol.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
Both methods stain connective tissue, but Masson–Goldner staining allows for the visualization and differentiation of muscles and connective tissue (connective tissue is green, muscles are brick red), whereas van Gieson staining differentiates collagen and elastin fibers (elastic fibers are black, and collagen fibers are red).
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
PHLDA1, ABCB1, cleaved PARP-1 and cleaved caspase-3 immunohistochemical reactions were performed via standard formalin-fixed, paraffin-embedded (FFPE) 4 μm slices of neuroblastoma tumors.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
ABCB1 antigens were retrieved using low-pH buffer (H-3300, Antigen Unmasking Solution, Citrate-based, pH 6.0; Vector Laboratories).
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
Endogenous peroxidase activity was blocked with 3% H2O2 solution.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
Membrane permeabilization was achieved with 0.1% Triton X-100 reagent.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
The tissue sections were incubated overnight with a 1:100 primary antibody mixture (see Table 1) at 4 °C.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
The next day, the sections were incubated for 30 min with secondary anti-rabbit antibodies (MP-7451, Vector Laboratories).
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
As positive controls, mouse liver and kidney sections were used.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
PHLDA1, cleaved PARP-1 and cleaved caspase-3 antigens were retrieved using high-pH buffer (H-3301, Antigen Unmasking Solution, Tris-based, pH 9.0; Vector Laboratories).
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
Endogenous peroxidase activity was blocked with 3% H2O2 solution.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
The samples were incubated overnight with a 1:40 (PHLDA1), 1:75 (cleaved PARP-1) or 1:50 (cleaved caspase-3) primary antibody mixture (see Table 1).
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
For PHLDA1 and cleaved PARP-1, the sections were incubated overnight at 4 °C, and for cleaved caspase-3, they were incubated for 3 h at room temperature.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
Then, the sections were incubated for 30 min with secondary anti-rabbit antibodies (MP-7451, Vector Laboratories).
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
For PHLDA1, positive controls for immunostaining of mouse kidney sections were used.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
The neuroblastoma with the greatest number of morphologically detected apoptotic cells served as a positive control for cleaved PARP-1 and cleaved caspase-3.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
The ImmPACT Nova Red (SK-4805, Vector Laboratories) reagent was used to visualize the immunostaining reaction.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
Nuclei were counterstained with Mayer’s hematoxylin.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
For PHLDA1 and ABCB1 staining the area of the tumors was examined in five random fields of vision for each group and, the both staining intensity, evaluated as 0-negative, 1-weak, 2-moderate, 3-strong.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
Then the percentage of IHC-positive cells was assessed and used for the staining index calculation according to the formula: [% x staining intensity].
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
Apoptosis in hematoxylin and eosin-stained samples was identified based on morphology.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
The number of cells with pyknotic nuclei and apoptotic bodies was counted under a bright-field microscope (Olympus Optical Co., Tokyo, Japan) with a 40x objective.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
Five adjacent fields of view present on each slide were examined.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
Apoptotic cells were also detected with an anti-cleaved PARP-1 antibody and an anti-cleaved caspase-3 antibody, and the number of cells with a positive IHC signal was counted with a 40x objective.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
The results of the counting of apoptotic cells are shown in the Supplementary Table S2a-f online.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
Bioinformatic screening was performed via the R2 platform (http://r2.amc.nl, http://r2platform.com).
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
The Versteeg 88 dataset was used for the analyses.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
Each of the in vitro experiments was conducted at least three times.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
The results are presented as the means (± SEM) of independent experiments.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
Statistical analyses of experiments involving two groups were performed via two-tailed Student’s t test for independent samples.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
Other experiments were statistically analyzed via the Kruskal‒Wallis ANOVA test, followed by Dunn’s post hoc analysis or one-way ANOVA followed by the post hoc Tukey’s test.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
Calculations were performed via Excel software (Microsoft), R (R version 3.2.1 Patched), and Graph Pad Prism 6.
PMC12359789
433. HTR2A EXPRESSION LEVELS ARE CORRELATED WITH THE RELEASE OF NEUROFILAMENT LIGHT CHAIN
In our previous study on Admission Delirium Tremens (ADT), a severe form of alcohol withdrawal marked by tremors and confusion, we found a significant increase in the neuronal biomarker neurofilament light chain (NfL) .
PMC12359789
433. HTR2A EXPRESSION LEVELS ARE CORRELATED WITH THE RELEASE OF NEUROFILAMENT LIGHT CHAIN
Although haloperidol, used for DT by antagonizing dopamine D2 and serotonin 5-HT2A receptors .,
PMC12359789
433. HTR2A EXPRESSION LEVELS ARE CORRELATED WITH THE RELEASE OF NEUROFILAMENT LIGHT CHAIN
was once a common treatment, it did not impact the duration of DT and is no longer the primary therapy.
PMC12359789
433. HTR2A EXPRESSION LEVELS ARE CORRELATED WITH THE RELEASE OF NEUROFILAMENT LIGHT CHAIN
Genetic association analyses revealed a correlation between the serotonin 5-HT2A receptor gene (HTR2A) and NfL. This study aims to investigate whether the expression level of HTR2A correlates with the release of NfL in cell culture media.
PMC12359789
433. HTR2A EXPRESSION LEVELS ARE CORRELATED WITH THE RELEASE OF NEUROFILAMENT LIGHT CHAIN
A total of 221 alcohol-dependent patients were enrolled for genotyping verification.
PMC12359789
433. HTR2A EXPRESSION LEVELS ARE CORRELATED WITH THE RELEASE OF NEUROFILAMENT LIGHT CHAIN
Plasma and culture media NfL levels were measured using the Simple Plex™ NfL Assay on the Ella™ instrument, following the manufacturer's instructions.
PMC12359789
433. HTR2A EXPRESSION LEVELS ARE CORRELATED WITH THE RELEASE OF NEUROFILAMENT LIGHT CHAIN
HTR2A SNPs were selected from the imputed dataset generated by the Michigan Imputation Server, and the polymorphism rs9567746 was validated using TaqMan Real-Time PCR assays.
PMC12359789
433. HTR2A EXPRESSION LEVELS ARE CORRELATED WITH THE RELEASE OF NEUROFILAMENT LIGHT CHAIN
Its association with NfL levels was then evaluated through linear regression analysis.
PMC12359789
433. HTR2A EXPRESSION LEVELS ARE CORRELATED WITH THE RELEASE OF NEUROFILAMENT LIGHT CHAIN
The SK-N-SH human neuroblastoma cell line was cultured, and HTR2A expression was knocked down using HTR2A siRNA targeting exon 4 (1466–1486, NM_000621.5) in the presence or absence of TNF-α or haloperidol.
PMC12359789
433. HTR2A EXPRESSION LEVELS ARE CORRELATED WITH THE RELEASE OF NEUROFILAMENT LIGHT CHAIN
HTR2A mRNA expression levels were quantified by real-time PCR.
PMC12359789
433. HTR2A EXPRESSION LEVELS ARE CORRELATED WITH THE RELEASE OF NEUROFILAMENT LIGHT CHAIN
Data visualization was conducted using GraphPad Prism 5 software, with statistical significance set at P < 0.05.
PMC12359789
433. HTR2A EXPRESSION LEVELS ARE CORRELATED WITH THE RELEASE OF NEUROFILAMENT LIGHT CHAIN
ADT patients exhibited significantly higher plasma NfL levels compared to non-DT patients and controls.
PMC12359789
433. HTR2A EXPRESSION LEVELS ARE CORRELATED WITH THE RELEASE OF NEUROFILAMENT LIGHT CHAIN
Notably, DT patients’ HTR2A genetic polymorphisms located at intron 3 were significantly associated with NfL levels (P = 0.001).
PMC12359789
433. HTR2A EXPRESSION LEVELS ARE CORRELATED WITH THE RELEASE OF NEUROFILAMENT LIGHT CHAIN
Carriers of the minor allele at rs9567746 showed significantly elevated NfL levels than the major genotype carriers.
PMC12359789
433. HTR2A EXPRESSION LEVELS ARE CORRELATED WITH THE RELEASE OF NEUROFILAMENT LIGHT CHAIN
In vitro, selective knockdown of HTR2A significantly increased NfL release in SK-N-SH cell culture media, particularly when co-incubated with TNF-α (100 ng/mL).
PMC12359789
433. HTR2A EXPRESSION LEVELS ARE CORRELATED WITH THE RELEASE OF NEUROFILAMENT LIGHT CHAIN
These findings suggest a mechanistic link between HTR2A expression and NfL release.
PMC12359789
433. HTR2A EXPRESSION LEVELS ARE CORRELATED WITH THE RELEASE OF NEUROFILAMENT LIGHT CHAIN
In rs9567746 polymorphism of HTR2A, the ADT patient’s minor allele type carrier, present in approximately 20% of ADT patients, showed significant increase of plasma NfL levels than major genotype carriers.
PMC12359789
433. HTR2A EXPRESSION LEVELS ARE CORRELATED WITH THE RELEASE OF NEUROFILAMENT LIGHT CHAIN
This minor genotype carriers could be having lower HTR2A expression, this could be demonstrated by HTR2A knock-down and increase the NfL release through pathways with or without TNF-α, highlighting the low expression of HTR2A could be a pathway toward more severe brain neuronal injury in ADT.
PMC12732662
Pharmacological Inhibition of JNK Signalling Exerts Anti-Neoplastic Effects on SH-SY5Y Human Neuroblastoma Cells
Neuroblastoma (NB) is the most prevalent paediatric extracranial solid tumour, which remains a major therapeutic challenge, especially in cases of recurrent and disseminated disease.
PMC12732662
Pharmacological Inhibition of JNK Signalling Exerts Anti-Neoplastic Effects on SH-SY5Y Human Neuroblastoma Cells
c-Jun N-terminal kinases (JNKs) are increasingly evidenced to play a key role in NB tumourigenesis and progression through apoptosis regulation, making selective JNK inhibitors promising candidates for use in targeted anticancer drugs in NB.
PMC12732662
Pharmacological Inhibition of JNK Signalling Exerts Anti-Neoplastic Effects on SH-SY5Y Human Neuroblastoma Cells
Our study comprehensively investigated the acute antineoplastic potential of the selective JNK inhibitor AS601245 (JNK inhibitor V) on the human MYCN-non-amplified neuroblastoma cell line, SH-SY5Y, with particular focus on its effects on NB cell viability, proliferation, migration, apoptosis, gene and protein expression, and mitochondrial metabolism.
PMC12732662
Pharmacological Inhibition of JNK Signalling Exerts Anti-Neoplastic Effects on SH-SY5Y Human Neuroblastoma Cells
JNK V selectively impaired NB cell survival and function, without exerting cytotoxicity toward normal human Schwann cells (HSC) and fibroblasts (BJ).
PMC12732662
Pharmacological Inhibition of JNK Signalling Exerts Anti-Neoplastic Effects on SH-SY5Y Human Neuroblastoma Cells
Our findings highlighted a dose-dependent inhibition of proliferation (XTT assay), colony formation (clonogenic assay), and migration (wound healing assay), accompanied by increased caspase-3 activity (caspase-3 assay), pro-apoptotic genes (qRT-PCR) and protein (Western blotting) expression, and significant disruption of both oxidative phosphorylation and glycolysis (Agilent Seahorse XF Assay).
PMC12732662
Pharmacological Inhibition of JNK Signalling Exerts Anti-Neoplastic Effects on SH-SY5Y Human Neuroblastoma Cells
These results provide new insights into the therapeutic potential of JNK inhibition as a targeted strategy for NB.Neuroblastoma (NB) is the most frequently occurring paediatric extracranial solid tumour, constituting up to 15% of all cancer deaths in children .
PMC12732662
Pharmacological Inhibition of JNK Signalling Exerts Anti-Neoplastic Effects on SH-SY5Y Human Neuroblastoma Cells
NB is an embryonal neuroendocrine neoplasm of the sympathetic nervous system, originating from neural crest progenitor cells, which undergo maladaptive differentiation because of genomic and epigenetic defects .