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Objective: This study aimed to evaluate the anticancer effect of NCB-0846, a TNIK inhibitor, in MCF-7 cells and to assess its impact on the expression levels of NF-κB and TNFA at the gene level. Materials and Methods: The MCF-7 cell line was cultured at 37°C in a 5% CO2 atmosphere using Dulbecco's Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and antibiotics (50 IU/mL penicillin and 50 mg/mL streptomycin). Cell viability was analyzed using the CCK-8 assay to determine the cytotoxic effect of NCB-0846. Acridine Orange/Propidium Iodide (AO/PI) staining was performed to evaluate the effect of NCB-0846 on cellular morphology in the MCF-7 cell line. Total RNA was isolated from cells treated with NCB-0846. Data analysis was performed using the SPSS 22.0 statistical program. Results: The data indicated that NCB-0846 significantly decreased the viability rates of MCF-7 cells in a dose-dependent manner (1-3 µM, p<0.01). RT-PCR analysis revealed that the expression level of NFKB1 increased 5.4-fold compared to the control group in MCF-7 cells treated with NCB-0846 at the effective dose and duration (p<0.01). In contrast, the expression level of TNFA decreased to 0.4-fold compared to the control group (p<0.01). Conclusion: The results demonstrate that NCB-0846 induces changes in the mRNA levels of the NFKB1 and TNFA genes, which are associated with inflammatory signalling pathways in MCF-7 cells. However, further molecular analyses are necessary to clarify the effect of NCB-0846 on inflammation in breast cancer and other cancer types. [ABSTRACT FROM AUTHOR]

Objective: This study aimed to evaluate the anticancer effect of NCB-0846, a TNIK inhibitor, in MCF-7 cells and to assess its impact on the expression levels of NF-κB and TNFA at the gene level. Materials and Methods: The MCF-7 cell line was cultured at 37°C in a 5% CO2 atmosphere using Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and antibiotics (50 IU/mL penicillin and 50 mg/mL streptomycin). Cell viability was analyzed using the CCK-8 assay to determine the cytotoxic effect of NCB- 0846. Acridine Orange/Propidium Iodide (AO/PI) staining was performed to evaluate the effect of NCB-0846 on cellular morphology in the MCF-7 cell line. Total RNA was isolated from cells treated with NCB-0846. Data analysis was performed using the SPSS 22.0 statistical program. Results: The data indicated that NCB-0846 significantly decreased the viability rates of MCF-7 cells in a dose-dependent manner (1-3 μM, p

Epilepsy, one of the most common neurological disorders, is characterized by spontaneous recurrent seizures. Temporal lobe epilepsy (TLE) is one of the most common medically intractable seizure disorders. Traf2-and NcK-interacting kinase (TNIK) has recently attracted attention as a critical modulation target of many neurological and psychiatric disorders, but its role in epilepsy remains unclear. In this study, we hypothesized the involvement of TNIK in epilepsy and investigated TNIK expression in patients with intractable TLE and in a pilocarpine-induced rat model of epilepsy by western blotting, immunofluorescence, and immunohistochemistry. A pentylenetetrazole (PTZ)-induced epilepsy rat model was used to determine the effect of the TNIK inhibitor NCB-0846 on behavioral manifestations of epilepsy. Coimmunoprecipitation (Co-IP)/mass spectrometry (MS) was used to identify the potential mechanism. Through Co-IP, we detected and confirmed the main potential TNIK interactors. Subcellular fractionation was used to establish the effect of NCB-0846 on the expression of the main interactors in postsynaptic density (PSD) fractions. We found that TNIK was primarily located in neurons and decreased significantly in epilepsy model rats and TLE patients compared with controls. NCB-0846 delayed kindling progression and decreased seizure severity. Co-IP/MS identified 63 candidate TNIK interactors in rat hippocampi, notably CaMKII. Co-IP showed that TNIK might correlate with endogenous GRIA1, SYN2, PSD-95, CaMKIV, GABRG1, and GABRG2. In addition, the significant decrease in GRIA1 in hippocampal total lysate and PSDs after NCB-0846 treatment might help modify the progression of PTZ kindling. Our results suggest that TNIK contributes to epileptic pathology and is a potential antiepileptic drug target.

Background: The treatment of patients with metastatic synovial sarcoma is still challenging, and the development of new molecular therapeutics is desirable. Dysregulation of Wnt signaling has been implicated in synovial sarcoma. Traf2-and-Nck-interacting kinase (TNIK) is an essential transcriptional co-regulator of Wnt target genes. We examined the efficacy of a small interfering RNA (siRNA) to TNIK and a small-molecule TNIK inhibitor, NCB-0846, for synovial sarcoma. Methods: The expression of TNIK was determined in 20 clinical samples of synovial sarcoma. The efficacy of NCB-0846 was evaluated in four synovial sarcoma cell lines and a mouse xenograft model. Results: We found that synovial sarcoma cell lines with Wnt activation were highly dependent upon the expression of TNIK for proliferation and survival. NCB-0846 induced apoptotic cell death in synovial sarcoma cells through blocking of Wnt target genes including MYC, and oral administration of NCB-846 induced regression of xenografts established by inoculation of synovial sarcoma cells. Discussion: It has become evident that activation of Wnt signaling is causatively involved in the pathogenesis of synovial sarcoma, but no molecular therapeutics targeting the pathway have been approved. This study revealed for the first time the therapeutic potential of TNIK inhibition in synovial sarcoma. [ABSTRACT FROM AUTHOR]