Purpose: Although chemotherapy is one of the standard treatments for gastric cancer, the disease’s resistance mechanisms continue to limit the survival rates. B7H6 (NCR3LG1), an immune checkpoint belonging to the B7 family, is significantly overexpressed in gastric cancer. This work investigated the possibility of using B7H6 suppression to improve the effectiveness of the widely used chemotherapy medication docetaxel. Materials and Methods: In this study, MKN-45 gastric cancer cells were transfected for 24 h with siRNA targeting B7H6, and then, docetaxel was added at optimal inhibitory doses (IC25 and IC50). To assess the impact of this combination therapy, cellular viability, proliferation, and migration were assessed using MTT assay, colony-forming unit assay, and wound-healing assay, respectively. Additionally, apoptosis and cell cycle status were evaluated by flow cytometry. Moreover, using qRT-PCR, the gene expression of B7H6 and indicators associated with apoptosis was also examined. Results: The sensitivity of MKN-45 cells to docetaxel was greatly increased by the siRNA-mediated knockdown of B7H6, resulting in a decrease in the drug’s IC50 value. When compared to each therapy alone, the combination of B7H6 siRNA plus docetaxel at IC50 levels exhibited a significant increase in apoptosis rate. The volume of cells arrested at the sub-G1 and G2-M phase was shown to rise when B7H6 siRNA transfection was combined with docetaxel. Furthermore, the combination treatment significantly decreased the ability of cells to migrate and form colonies. Conclusions: B7H6 suppression increases the susceptibility of MKN-45 gastric cancer cells to docetaxel treatment, resulting in decreased cellular proliferation and increased rates of apoptosis. The present work underscores the possibility of enhancing treatment results in gastric cancer by merging conventional chemotherapy with gene-silencing approaches.

Background. Pancreatic ductal adenocarcinoma remains difficult to treat and has an exceptionally poor prognosis. While prior studies have noted antitumour properties for midazolam in pancreatic cancer, whether this response involves developmental signaling pathways is unclear. Here, we evaluated midazolam-treated MIA PaCa-2 cells to identify associated Wnt and Hedgehog transcriptional shifts and their systems-level context. Methods. MIA PaCa-2 cells were exposed to midazolam (20–100 µM) for 24 h. We measured metabolic viability by MTT and apoptosis by Annexin V-FITC/7-AAD flow cytometry at 60 µM. Expression of 18 targeted genes was quantified by RT-qPCR using ACTB for normalization. Changed genes were examined through STRING/KEGG networks, TCGA-PAAD survival analysis, DGIdb annotations, and preliminary FZD7 molecular docking. Results. Midazolam reduced MTT-derived metabolic viability in a concentration-dependent manner (IC₅₀ = 54.6 µM) and significantly increased Annexin V-positive apoptotic populations at 60 µM across three independent experiments. Eight transcripts changed significantly: CTNNB1, TGFBR2, WNT2, WNT7B, HHIP, and DHH declined, while CSNK1A1 and AXIN1 increased (all adjusted p < 0.05). Network analysis highlighted CTNNB1 as the main hub, and high CTNNB1 expression correlated with shorter survival in TCGA-PAAD patients (p = 0.043; HR = 1.58, 95% CI: 1.01–2.47). Conclusions. Midazolam reduced metabolic viability and increased apoptosis in MIA PaCa-2 cells alongside coordinated expression changes in key Wnt and Hedgehog components. Because the required doses far exceed clinical sedation levels and pathway activity was not directly confirmed at the protein level, these findings should be regarded as a preliminary hypothesis rather than evidence of therapeutic efficacy.

Marinesco–Sjögren syndrome (MSS) is a rare autosomal recessive disorder characterized by cerebellar ataxia, congenital cataracts, developmental delay, hypotonia, and progressive myopathy. Most reported cases are linked to pathogenic variants in SIL1, a gene encoding a co-chaperone essential for protein folding in the endoplasmic reticulum. Here, we present a comprehensive case study of a Turkish pediatric patient diagnosed with MSS, supported by genetic, bioinformatic, and structural modeling analyses. Whole-exome sequencing revealed a homozygous splice-site variant (SIL1 c.453+1G>T), confirmed by Sanger sequencing and segregation analysis. In silico annotation using Genomize, InterVar, Franklin, VarSome, ClinVar, OMIM, and PubMed classified the variant as pathogenic according to ACMG guidelines. Structural modeling by Phyre2 and I-TASSER demonstrated that the variant abolishes the intron 5 donor site, leading to truncation of the wild-type 461-amino-acid protein into a shortened ~189-amino-acid polypeptide. This truncation results in the loss of critical Armadillo (ARM) repeats required for HSPA5 interaction, explaining the observed instability and impaired chaperone function. Clinically, the patient presented with congenital cataracts, ataxia, developmental delay, and progressive muscle weakness, consistent with previously reported MSS cases. Comparison with the literature confirmed that splice-site variants frequently correlate with severe phenotypes, including early-onset ataxia and cataracts. This report highlights the importance of integrating genomic, structural, and clinical data to better understand genotype–phenotype correlations in MSS. Our findings expand the mutational spectrum of SIL1, reinforce the role of splicing defects in disease pathogenesis, and emphasize the necessity of comprehensive molecular diagnostics for rare neurogenetic syndromes.

Pancreatic ductal adenocarcinoma (PDAC) remains a highly lethal malignancy, and there is continuing interest in whether agents used during the perioperative period influence tumour-cell biology. Propofol has been associated with antiproliferative and pro-apoptotic effects in experimental cancer models; however, its relationship with Wnt–Frizzled-related transcriptional responses in pancreatic cancer cells remains incompletely characterised. MIA PaCa-2 cells were exposed to propofol for 24 h. Cellular metabolic activity was assessed using the MTT assay over a concentration range of 0–20 µM, and apoptosis was evaluated via Annexin V-FITC/7-AAD flow cytometry at the MTT-derived IC₅₀ concentration of 7.6 µM. Transcript abundance of 18 pathway-associated genes was analysed via RT-qPCR following exposure to 7.6 µM propofol. MTT, flow-cytometry, and RT-qPCR assays were each performed in three independent biological experiments (n = 3). RT-qPCR results were evaluated after Benjamini–Hochberg false discovery rate correction across the 18-transcript panel. Complementary bioinformatic analyses were performed solely to provide exploratory biological and clinical context for the experimentally prioritised transcripts. Propofol produced a concentration-dependent reduction in normalised MTT metabolic activity, with an MTT-derived IC₅₀ of 7.6 µM. At this concentration, total apoptosis increased from 0.67 ± 0.11% in vehicle-treated cells to 17.04 ± 2.40% following propofol exposure (p < 0.001), whereas necrosis remained minimal. The viable-cell fraction decreased from 99.30 ± 1.057% to 82.80 ± 0.841% (p < 0.05). The difference between the MTT-derived IC₅₀ and the Annexin V/7-AAD viable-cell fraction indicates that the MTT-derived IC₅₀ reflects reduced metabolic activity rather than 50% cell lethality. RT-qPCR showed decreased WNT5B, FZD5, and WNT11 transcript abundance and increased GSK3B transcript abundance. All four remained significant after Benjamini–Hochberg correction (WNT5B, q = 0.001; FZD5, q = 0.001; WNT11, q = 0.027; GSK3B, q = 0.041). Exploratory bioinformatic analyses showed heterogeneous gene-specific expression, survival, genomic, immune-infiltration, and dependency patterns and were not interpreted as evidence of a propofol-mediated mechanism in patients. Under the tested in vitro conditions, propofol reduced MTT-derived metabolic activity, increased apoptosis, and was associated with selective Wnt–Frizzled-related transcript changes in MIA PaCa-2 cells. These transcript-level observations do not establish direct functional modulation of Wnt–Frizzled signalling or demonstrate that this pathway mediates the observed apoptotic response. The bioinformatic findings provide exploratory contextual information only. Further validation in additional PDAC and non-malignant pancreatic cell models, together with protein-, phosphorylation-, and pathway-level functional studies, is required.