Anthocyanins from red grapes possess notable anticancer properties, yet their biological activity after gastrointestinal digestion remains poorly characterized. In this study, we investigated the in vitro antitumor effects of a red grape anthocyanin extract digested using the INFOGEST protocol on colorectal cancer (CRC) cell lines. The study assessed cell viability, apoptosis induction, cell cycle progression, and confined 3D migration, and integrated whole-transcriptome microarray profiling to elucidate the underlying molecular mechanisms.Cell viability assays showed that simulated digestion markedly increased the extract’s potency, reducing the IC50 values by 2.25-fold in DLD1 cells and 1.59-fold in Caco2 cells. Transcriptomic profiling by microarray analysis demonstrated substantial gene expression changes upon treatment in DLD1 cells, with 873 genes showing a fold regulation greater than |±1.5|. Gene Set Enrichment Analysis (GSEA) identified enrichment of pro-apoptotic gene sets in treated cells, including p53 signaling, prolonged unfolded protein response activation and suppression of RNA metabolism. These findings were supported by the Annexin V/PI assay demonstrating apoptosis induction, and RT-qPCR validation of key apoptosis-regulating genes. The extract also displayed significant antiproliferative activity with cells arrested in the S phase, as evidenced by AlamarBlue and cell cycle analyses. Downregulation of key cell cycle promoters and upregulation of proliferation-inhibitory genes were also observed. Furthermore, the extract significantly impaired 3D migration in a microfluidic model, reducing migration speed, directionality, and number of migratory cells. GSEA revealed suppression of RHO GTPases and MIRO-mediated signaling, suggesting disruption of cytoskeletal and mitochondrial dynamics, strongly impacting cell migration. Together, these findings indicate that digested red grape anthocyanins exert a multifaceted anticancer effect in vitro by promoting apoptosis, inhibiting proliferation, and impairing cell migration, highlighting their potential as functional bioactive compounds for CRC management.

Background: Germline multigene testing is increasingly integrated into breast cancer care, but the ability of current eligibility frameworks to identify pathogenic variant carriers remains uncertain in real-world cohorts. Methods: This retrospective observational study included 556 patients with histologically confirmed breast cancer who underwent germline multigene testing at a single oncology center. Testing eligibility was retrospectively assessed according to NCCN criteria, ESMO-based recommendations, and the ASBrS universal-testing recommendations. Family history up to third-degree relatives and surgical data were evaluated. Results: Pathogenic variants were identified in 137 patients (24.6%), while variants of uncertain significance were reported in 310 patients (55.8%). NCCN criteria were fulfilled by 135 of 137 patients with pathogenic variants (98.5%), indicating high sensitivity but limited discriminatory capacity, as eligibility was also frequent among patients without pathogenic variants. ESMO-based criteria were met by 94 mutation-positive patients (68.6%) and were significantly associated with pathogenic variant status (p < 0.001; OR = 3.08, 95% CI: 2.05-4.65). Notably, 42.3% of pathogenic-variant-positive patients reported no family history of malignancy. Prophylactic surgery was documented in only 16.8% of patients with pathogenic variants. Conclusions: NCCN criteria captured nearly all mutation-positive patients; ESMO-based criteria were more selective. The findings support broader access to germline testing before surgery.

Abstract Metastasis is the leading cause of mortality in breast cancer and remains largely untargeted therapeutically. Identifying molecular drivers of metastatic progression is essential for developing effective treatments. This study investigated the role of the transcription factor ELK3 in triple-negative breast cancer (TNBC) metastasis by defining the cellular and molecular processes it regulates. MDA231 cells with ELK3 overexpression (OE) or knockdown (KD) were generated by lentiviral transduction. Transcriptomic alterations induced by ELK3-KD were analyzed by microarray and validated by RT-qPCR. Ingenuity Pathway Analysis and Gene Set Enrichment Analysis identified ELK3-dependent metastasis-associated pathways, which were functionally validated using 3D microfluidic migration assays, mammosphere formation assays, and flow cytometry/ AlamarBlue proliferation assays. High ELK3 expression correlated with a mesenchymal phenotype in BC cell lines and lymph node invasion in patient tumors. ELK3-KD significantly altered 740 genes, many linked to migration and stemness. Functionally, ELK3 enhanced 3D confined migration, likely through regulation of EMT, cell adhesion and protrusion formation. ELK3 also promoted cancer stem cell traits, potentially via hypoxia-related and WNT/β-catenin, JAK/STAT3, TGF-β, Notch1, and NF-κB signaling pathways. Additionally, ELK3 induced cellular quiescence while suppressing proliferation under adherent conditions. Overall, ELK3 acts as a pro-metastatic regulator in TNBC by promoting migration and stemness.

Background: Conventional axillary ultrasound (US) is least reliable in lymph nodes with diffusely thickened cortex. We evaluated whether contrast-enhanced ultrasound with ultra-resolution microvascular imaging (CEUS-URM) improves discrimination of metastatic nodes in this subgroup. Materials and methods: This was a prospective single-center study of patients with histologically confirmed breast cancer; one index (most suspicious) node per patient (unit of analysis = index node). Two separately recruited consecutive cohorts were analyzed: a US-only cohort (n = 181; diffuse-thickening subgroup with histology, n = 52) and a subsequent CEUS-URM cohort (n = 42; 15 metastatic). Surgical histopathology (SLNB/ALND) was the reference standard. A CEUS-URM score was built from data-driven, Youden-optimized cut-offs (URM vessel count ≥8; DV mean density ≥ 13.05) and internally validated by bootstrap with optimism correction. As the cohorts were separate, the US-versus-CEUS comparison is cross-cohort and exploratory. Results: Within the diagnostically challenging subgroup of lymph nodes with diffusely thickened cortex, conventional axillary US alone demonstrated limited discriminatory performance for metastatic involvement (AUC 0.43). CEUS-derived quantitative parameters significantly improved diagnostic accuracy, with the best individual parameter achieving an AUC of 0.68. A simple CEUS score combining hypervascular vessel count and vascular density provided the highest diagnostic performance (AUC 0.81, 95% CI 0.68-0.92). At a low threshold, the CEUS score showed high sensitivity (93%), suitable for screening and exclusion of nodal metastases, while at a higher threshold it achieved high specificity (96%), allowing reliable confirmation of metastatic disease. Conclusions: In this exploratory study, a simple CEUS-URM score improved discrimination of diffusely thickened axillary nodes and may serve as an adjunct to conventional US. The findings are preliminary-derived and tested in the same small cohort-and require external, within-patient paired validation.

BACKGROUND:Early detection is critical in pancreatic ductal adenocarcinoma (PDAC)-one of the most lethal malignancies due to its typically late diagnosis. METHODS:In this study, we aimed to validate an epidemiological risk score (ERS) designed to identify individuals at increased risk of developing PDAC prior to the use of imaging or other diagnostic procedures. The ERS was constructed through a meta-analysis of 24 well-established epidemiological risk factors. We applied this score to a prospective cohort of 178 high-risk individuals with a family history of PDAC within the IMAGene project (ClinicalTrials.gov registration code: NCT06334458). To evaluate the predictive value of the ERS, all participants underwent whole-body or abdominal magnetic resonance imaging (MRI) and the findings were classified according to the Oncologically Relevant Findings Reporting and Data System criteria to identify and categorize lesions based on their malignant potential. External validation was conducted by using a subset of the UK Biobank (UKB) cohort (≈300 000 individuals), among whom 1648 were diagnosed with PDAC. RESULTS:Higher ERS values were associated with the presence of potentially malignant lesions on MRI. Both pancreatic and extra-pancreatic malignant lesions were more frequent among individuals with higher ERS scores (P = .01 and P = .02 respectively) compared with controls. External validation in PDAC cases within the UKB cohort confirmed these associations. CONCLUSION:Our findings support the integration of the ERS as a feasible, low-cost tool for PDAC risk stratification, with the potential to facilitate earlier detection and improve clinical outcomes.

Background and Objectives: Over recent decades, germline genetic testing has become increasingly integrated into breast cancer care, yet its precise effect on the timing of surgical workflows remains incompletely defined. This study investigates the implementation of nationally reimbursed genetic testing programs and examines how the scheduling of multigene germline testing relates to surgical timing. Materials and Methods: In this retrospective cohort analysis, we examined 502 breast cancer cases from the institutional registry of the "Ion Chiricuță" Oncology Institute (IOCN), with a mean age of 52 years, a high rate of neoadjuvant therapy (82.3%) and a majority of them in T2 (52%) and N1 (43%), N2 (29%) clinical stage: 263 patients from an earlier private-practice pay testing era (pre-reimbursement) as a comparison sample and 239 patients from the period when reimbursement programs were in operation. We then evaluated the benefits of state-funded genetic testing initiatives, computing three intervals: diagnosis to genetic test, genetic test to surgery, and diagnosis to surgery. Patients were categorized by timing of multigene testing (preoperative vs. postoperative), receipt of neoadjuvant systemic therapy (NACT), mutation status, and funding source for testing (national PNS program funded by the Romanian Ministry of Health; PNRR European Recovery and Resilience Facility funds; or self-funded private testing). Nonparametric statistics (Mann-Whitney U, Spearman correlation) and effect-size metrics (Cliff's delta, Theil-Sen slope) were employed. Results: The median diagnosis-to-surgery interval was 203 days (IQR 179-230). Patients tested preoperatively had longer intervals than those tested postoperatively (216 vs. 182.5 days; p = 0.000153; Cliff's δ = -0.486), a pattern driven by shared pathways involving NACT rather than by testing-induced delays. NACT was the principal determinant of surgical timing (211 vs. 43 days; p = 302.55 × 10-11). Within the preoperative subgroup, time to multigene testing correlated strongly with time to surgery (Spearman ρ = 0.54; p = 4.75 × 10-7; Theil-Sen slope = 0.37, 95% CI = 0.22-0.53). However, the no-NACT group included only four patients. The presence of a pathogenic variant did not significantly change surgical timing (p = 0.982). The PNS national reimbursement program achieved the highest preoperative integration rate for multigene genetic testing after adjustment for NACT confounder (76.7%), outperforming PNRR (61%) and private practice (56%). Conclusions: While genetic testing timing correlates with surgical workflow, neoadjuvant systemic therapy pathways and program structure exert greater influence than testing per se. Structured national programs enhance preoperative testing uptake without causing delays beyond those inherent to NACT pathways.

Background/Objectives: Molecular tumor profiling has recently transformed oncologic care delivery, establishing precision medicine as an essential approach for defining cancer biology and revealing intratumoral heterogeneity. The growing accessibility of advanced nucleic acid sequencing technologies has created a demand for specialized expertise in interpreting comprehensive genomic profiling results. Academic institutions currently employ a strategy of conducting initial broad-spectrum genomic testing, followed by matching patients to investigational therapies targeting their specific genomic alterations. Consequently, molecular tumor boards (MTBs) have emerged predominantly within major cancer centers and academic medical institutions, providing the specialized knowledge necessary to translate precision oncology into routine clinical care. However, despite the substantial benefits of collaborative case review within tumor boards, clinicians frequently encounter multiple barriers to effective MTB implementation. Methods: this report examines these challenges performing an exploratory quantitative synthesis approach and explores implementation strategies and best practices derived from collective institutional experiences, with the goal of establishing a functional MTB at the local level and thereby expanding oncology patient access to cutting-edge therapeutic options.

MiR-210 is widely recognized as the quintessential hypoxia-responsive miRNA and is thought to fine-tune various facets of cellular homeostasis. We hereby present an integrative appraisal of the phenotypic and molecular repercussions of disrupting the corresponding locus in human and mouse cells using multiple genetic strategies. In brief, MIR210 deletion led to decreased cellular fitness and suboptimal responses to several stress types. Transcriptomic comparisons via different profiling platforms, performed independently by members of this collaboration, revealed consistent deregulation of neighboring genes, in locus-disrupted cells. Interestingly, the anticipated enrichment of miR-210 targets failed to materialize in unbiased analyses. Our results point to the biological significance of unrecognized regulatory elements that overlap miRNA genes and should serve as a note of caution for studies based on the genetic disruption of such loci.

Triple-negative breast cancer (TNBC) is an aggressive subtype with high heterogeneity, metastasis, and drug resistance, underscoring the need for new therapeutic strategies. Sea buckthorn berries, long used in traditional medicine, are rich in carotenoids with reported anticancer activity. To investigate their transcriptional effects, we treated BT-549 TNBC cells with a saponified lipophilic sea buckthorn berry extract (LSBE) and performed gene expression profiling using microarray (four independent replicates of treated, respectively, untreated cells). LSBE altered the expression of 330 protein-coding genes (111 downregulated, 219 upregulated; fold ratio ±1.5, adj. p < 0.05). ingenuity pathway analysis (IPA) revealed activation of cholesterol biosynthesis (z = 3.46, p = 2.7E-15) and ferroptosis (z = 0.78, p = 5.3E-09), while oxidative stress (z = -2.18, p = 3.4E-05) and cholesterol accumulation (z = -2.14, p = 3.4E-06) were inhibited. Gene set enrichment analysis (GSEA) further indicated upregulation of cholesterol homeostasis and mTORC1 signaling. Our findings suggest that carotenoid-enriched LSBE modulates key pathways related to cellular homeostasis, antioxidant defense, and apoptosis in TNBC. While limited to transcriptional profiling, this exploratory study provides a foundation for future functional validation of LSBE as a complementary TNBC therapeutic approach.

Acquired resistance to chemotherapy, including irinotecan, remains a major challenge in treating metastatic colorectal cancer (CRC). Natural compounds such as curcumin have demonstrated potential in resensitizing chemoresistant cancer cells to existing therapies. This study investigates curcumin's ability to reverse irinotecan resistance in CRC cells and the underlying molecular mechanisms. An irinotecan-resistant CRC cell line (DLD1_IRI-R) was established by gradually increasing irinotecan exposure. The DLD1 cell line was selected for its intermediate sensitivity to irinotecan among CRC cell lines. Cell sensitivity to irinotecan and curcumin was assessed using the MTT assay, with drug interactions evaluated via the Chou-Talalay method. Apoptosis and cell cycle progression were analyzed by flow cytometry, proliferation by clonogenic assays, and migration in 3D microfluidic systems. Whole-genome transcription profiling was conducted using microarrays, with functional analysis performed in Ingenuity Pathway Analysis. DLD1_IRI-R cells exhibited a 7.17-fold increase in irinotecan resistance, accompanied by reduced proliferation and migration. Resistance acquisition led to dysregulation of genes involved in irinotecan metabolism (CYPs, UGTs, AKRs), efflux transport (ABCs), and ER stress adaptation. The gene coding for the drug target, TOP1, was also inhibited. Curcumin, combined with irinotecan at IC10, reduced irinotecan's IC50 by 3.74-fold, exhibiting strong synergy. Curcumin significantly modulated 3,901 genes (FC > |±2|), inducing apoptosis, disrupting ER stress adaptation, reducing proliferation, and inhibiting migration. It also upregulated TOP1 while suppressing key resistance-associated genes, including 18 CYPs, 4 UGTs, 4 AKRs, and 20 ABCs. These findings suggest that low-dose curcumin effectively reverses irinotecan resistance in CRC cells, enhancing chemotherapy sensitivity and inhibiting metastasis-associated traits.

Background and Objectives: The assessment of HER2 status in invasive breast carcinomas (IBCs) is critical for determining treatment strategies. The aim of this study was to evaluate the FICTION technique as a potential method for assessing HER2 status and to compare it with the standard sequential immunohistochemistry (IHC)-in situ hybridization (ISH) assays. Materials and Methods: This study included 49 patients diagnosed with invasive breast carcinomas. HER2 status was assessed using both IHC+FISH and FICTION techniques, and the results were compared. Results: Comparative analysis demonstrated an 83.67% categorical agreement between IHC and IF using the ASCO/CAP system. The percentage of cells showing any degree of HER2 protein expression was higher with IF (73.77%) than with IHC (60.71%) (p = 0.00026). The in situ hybridization assays showed an excellent agreement, with a 90% or higher concordance. The concordance of the ASCO/CAP group classification of cases using both ISH assays (FICTION and standard FISH) was high (85, 7%). Agreement was 100% for the final classification of cases (Her2 positive/negative). Conclusions: We compared standard tests for Her2 protein expression and the gene copy number with a modified FICTION protocol. The study showed moderate agreement between IHC and IF for Her2 protein and excellent agreement between FISH and FICTION ISH for the gene copy number. Final Her2 status was unaffected by low IF IHC concordance. Optimizing the FICTION protocol could improve results. Combining protein and gene assays may enhance IBC patient stratification.

OBJECTIVES:Interval debulking surgery has similar outcomes and less morbidity compared with primary debulking in advanced ovarian cancer. However, there is controversy regarding the selection of chemotherapy-resistant clones. Complete resection is an essential prerequisite, and near-infrared surgery combined with various techniques for highlighting malignant foci strives to achieve actual complete resection. This study investigated the role of indocyanine green (ICG) in identifying additional residual malignant foci during interval debulking of apparently intact peritoneum not deemed clinically suspicious under white light inspection. METHODS:Patients diagnosed with stage III or IV high-grade serous ovarian carcinoma, older than 18 years of age, with satisfactory hepatic and renal functions who underwent neoadjuvant chemotherapy according to the institutional protocol and were scheduled to undergo interval debulking surgery between 2020 and 2022 were deemed suitable for inclusion after agreeing to the study protocol and acknowledging no contraindications for the administration of the ICG product. After laparotomy and white light inspection, using bolus administration of ICG, additional suspect peritoneal samples in near infrared (defined by clinical hyper- or hypointensity areas compared with surrounding ICG fluorescence using the Zeiss Opmi Pentero 800 surgical microscope, that were not deemed clinically suspicious under white light) were excised. Descriptive statistics were inferred and the chi-square test was used for the comparison of excised areas. The Kaplan-Meier method was deployed for computing the overall survival and progression-free survival of the cohort. All statistical analyses were performed using IBM SPSS Statistics software. RESULTS:Fifteen patients with a median age of 56 years were included. Most cases (n=10, 66.7%) were International Federation of Gynecology and Obstetrics (FIGO) stage III, and all patients received four to seven cycles of neoadjuvant platinum chemotherapy, with 40% of regimens using bevacizumab. The mean interval between neoadjuvant treatment and surgery was 39 (median 42, range 20-78) days. A total of 39 suspect additional peritoneal samples were analyzed, with 41% confirming malignant foci. The positive predictive value (PPV) for malignant foci was 30% in ICG hyperintense areas and 46% in ICG hypointense areas. Germline BRCA1/2 mutant patients and using neoadjuvant bevacizumab led to a higher PPV for ICG hypointense areas (60% and 72.7%, respectively). Overall, the number of additionally resected pathologically confirmed malignant lesions through ICG fluorescence increased by 25%. CONCLUSIONS:The use of ICG was associated with an increase in the resection of samples with residual malignant foci. Overall, hypointense areas had a higher positive PPV for malignant foci in comparison with hyperintense ICG areas (46% vs 30%), which could be interpreted in the context of dynamic changes in the tumor microenvironment or enhanced permeability and retention effect following neoadjuvant chemotherapy.

New meso-substituted AB3-type phenothiazinyl porphyrins and ferrocenylvinyl phenothiazinyl porphyrin were synthesised by Suzuki-Miyaura and Mizoroki-Heck cross-coupling reactions, respectively. The free porphyrins were further used in the synthesis of new indium(iii) or zinc(ii) porphyrin complexes. All porphyrins exhibit red fluorescence emission in solution, a property that remains unimpaired following internalisation in ovarian A2780 cancer cells, as evidenced by fluorescence microscopy images. The In(iii) phenothiazinyl porphyrin complexes show a higher quantum yield of fluorescence emission (2a Phi F = 30%, 4a Phi F = 29%, 5a Phi F = 28%) compared to the free base porphyrin precursors, or Zn(ii) complex 4b (Phi F = 10%). The potential of novel phenothiazinyl porphyrins to act as photosensitisers was evaluated using two distinct approaches. The first was through the measurement of the singlet oxygen quantum yield Phi Delta(1O2), while the second employed in vitro measurements of metabolic activity, oxidative stress, nuclear factor-erythroid 2 related factor 2 (Nrf-2) activation and tumour necrosis factor-alpha (TNF-alpha) under both dark and light irradiation conditions. As reflected by the IC50 values, the most potent cytotoxicity of the phenothiazinyl porphyrins against the A2780 cells was observed for In(iii) ferrocenylvinyl phenothiazinyl porphyrin 4a (36.38 mu M), the remaining compounds are less cytotoxic. The reduction in metabolic activity was observed in A2780 ovarian tumour cells treated with 4a and 6a and exposed to light compared to treatment in the absence of light. The oxidative stress, TNF-alpha and Nrf-2 transcription factor were particularly notable when A2780 cells were treated with 4a and subsequently photoirradiated, the oxidative stress was linked to the highest value of Phi Delta(1O2) recorded for 4a (60%).

Background and aims. Breast cancer (BC) is the most frequently diagnosed cancer and the leading cause of cancer-related death among women worldwide. For locally advanced diseases and high-risk tumors, neoadjuvant therapy (NAT) is the treatment of choice. Some studies show that mammographic density (MD) tumor margins and the presence of microcalcifications play a prognostic role in BC patients. Hence, the objective of this retrospective study was to assess if MD could predict the response to NAT among different molecular subtypes of BC patients undergoing NAT at The “Prof. Dr I. Chiricuta” Oncology Institute of Cluj-Napoca, Romania (IOCN). Furthermore, the association between MD, tumor margins and the presence of microcalcifications with clinico-pathological data was analyzed. Methods. Eighty-four breast cancer patients diagnosed and treated at IOCN were included in this study. The morphological characteristics of the tumors were framed according to the BIRADS lexicon. The presence or absence of microcalcifications was also assessed. First, the significance of associations between breast density, margins and microcalcifications and clinico-pathological parameters of the patients were tested with Fisher or Fisher-Freeman-Halton Exact Test. Next, using multinomial logistic regression, we modelled the associations between the pathological response measured by Miller Payne and Residual cancer burden (RCB) systems and the BI-RADS. Variables having significant univariate tests were selected as candidates for the multivariable analysis (adjusted model). Results. Breast densities were significantly associated with the age of the patients (p=0.01), number of positive lymph nodes (p=0.037), margins (p=0.002) and combined categories of Miller-Payne (p=0.034) and RCB pathological response (p=0.021). Margins was significantly associated with ki67 proliferation index (p=0.029), estrogen receptor (ER) (p=0.007), progesterone receptor (PR) (p=0.019), molecular subtype (p<0.001) and the number of clinically observed positive lymph nodes at diagnosis (p=0.019). Conclusions. In our cohort, BC patients with lower MD had higher odds of achieving pCR following NAT, suggesting the role of MD as a clinical prognostic marker. Larger multicenter studies are warranted to validate the prognostic value of MD, which could aid in patients stratification based on their likelihood to respond to NAT

BACKGROUND:Cancer stem-like cells (CSCs) have been extensively researched as the primary drivers of therapy resistance and tumor relapse in patients with breast cancer. However, due to lack of specific molecular markers, increased phenotypic plasticity and no clear clinicopathological features, the assessment of CSCs presence and functionality in solid tumors is challenging. While several potential markers, such as CD24/CD44, have been proposed, the extent to which they truly represent the stem cell potential of tumors or merely provide static snapshots is still a subject of controversy. Recent studies have highlighted the crucial role of the tumor microenvironment (TME) in influencing the CSC phenotype in breast cancer. The interplay between the tumor and TME induces significant changes in the cancer cell phenotype, leading to the acquisition of CSC characteristics, therapeutic resistance, and metastatic spread. Simultaneously, CSCs actively shape their microenvironment by evading immune surveillance and attracting stromal cells that support tumor progression. METHODS:In this study, we associated in vitro mammosphere formation assays with bulk tumor microarray profiling and deconvolution algorithms to map CSC functionality and the microenvironmental landscape in a large cohort of 125 breast tumors. RESULTS:We found that the TME score was a significant factor associated with CSC functionality. CSC-rich tumors were characterized by an immune-suppressed TME, while tumors devoid of CSC potential exhibited high immune infiltration and activation of pathways involved in the immune response. Gene expression analysis revealed IFNG, CXCR5, CD40LG, TBX21 and IL2RG to be associated with the CSC phenotype and also displayed prognostic value for patients with breast cancer. CONCLUSION:These results suggest that the characterization of CSCs content and functionality in tumors can be used as an attractive strategy to fine-tune treatments and guide clinical decisions to improve patients therapy response.

Aims While enhanced tumor cell migration is a key process in the tumor dissemination, mechanistic insights into causal relationships between tumor cells and mechanical confinement are still limited. Here we combine the use of microfluidic platforms to characterize confined cell migration with genomic tools to systematically unravel the global signaling landscape associated with the migratory phenotype of breast cancer (BC) cells. Meterials and methods The spontaneous migration capacity of seven BC cell lines was evaluated in 3D microfluidic devices and their migration capacity was correlated with publicly available molecular signatures. The role of identified signaling pathways on regulating BC migration capacity was determined by receptor stimulation through ligand binding or inhibition through siRNA silencing. Downstream effects on cell migration were evaluated in microfluidic devices, while the molecular changes were monitored by RT-qPCR. Key findings Expression of 715 genes was correlated with BC cells migratory phenotype, revealing TNF-α as one of the top upstream regulators. Signal transduction experiments revealed that TNF-α stimulates the confined migration of triple negative, mesenchymal-like BC cells that are also characterized by high TNFR1 expression, but inhibits the migration of epithelial-like cells with low TNFR1 expression. TNFR1 was strongly associated with the migration capacity and triple-negative, mesenchymal phenotype. Downstream of TNF/TNFR1 signaling, transcriptional regulation of NFKB seems to be important in driving cell migration in confined spaces. Significance TNF-α/TNFR1 signaling axis reveals as a key player in driving BC cells confined migration, emerging as a promising therapeutic strategy in targeting dissemination and metastasis of triple negative, mesenchymal BC cells.

Currently, only a limited set of molecular traits are utilized to direct treatment for metastatic CRC (mCRC). The molecular classification of CRC depicts tumor heterogeneity based on gene expression patterns and aids in comprehending the biological characteristics of tumor formation, growth and prognosis. Additionally, it assists physicians in tailoring the therapeutic approach. Microsatellite instability (MSI-H)/deficient mismatch repair proteins (MMRd) status has become a ubiquitous biomarker in solid tumors, caused by mutations or methylation of genes and, in turn, the accumulation of mutations and antigens that subsequently induce an immune response. Immune checkpoint inhibitors (ICI) have recently received approval for the treatment of mCRC with MSI-H/MMRd status. However, certain individuals experience either initial or acquired resistance. The tumor-programmed cell death ligand 1 (PD-L1) has been linked to the ability of CRC to evade the immune system and promote its growth. Through comprehensive research conducted via the PUBMED database, the objectives of this paper were to review the molecular characteristics linked to tumor response in metastatic CRC in light of improved patients’ outcomes following ICI therapies as seen in clinical trials and to identify particular microRNAs that can modulate the expression of specific oncoproteins, such as PD-L1, and disrupt the mechanisms that allow the immune system to be evaded.

Crops are under constant pressure due to global warming, which unfolds at a much faster pace than their ability to adapt through evolution. Agronomic traits are linked to cytoplasmic-nuclear genome interactions. It thus becomes important to understand the influence exerted by the organelles on gene expression under heat stress conditions and profit from the available genetic diversity. Maize (Zea mays) cytolines allow us to investigate how the gene expression changes under heat stress conditions in three different cytoplasmic environments, but each having the same nucleus. Analyzing retrograde signaling in such an experimental set-up has never been done before. Here, we quantified the response of three cytolines to heat stress as differentially expressed genes (DEGs), and studied gene expression patterns in the context of existing polymorphism in their organellar genomes. Our study unveils a plethora of new genes and GO terms that are differentially expressed or enriched, respectively, in response to heat stress. We report 19,600 DEGs as responding to heat stress (out of 30,331 analyzed), which significantly enrich 164 GO biological processes, 30 GO molecular functions, and 83 GO cell components. Our approach allowed for the discovery of a significant number of DEGs and GO terms that are not common in the three cytolines and could therefore be linked to retrograde signaling. Filtering for DEGs with a fold regulation > 2 (absolute values) that are exclusive to just one of the cytolines, we find a total of 391 up- and down-DEGs. Similarly, there are 19 GO terms with a fold enrichment > 2 that are cytoline-specific. Using GBS data we report contrasting differences in the number of DEGs and GO terms in each cytoline, which correlate with the genetic distances between the mitochondrial genomes (but not chloroplast) and the original nuclei of the cytolines, respectively. The experimental design used here adds a new facet to the paradigm used to explain how gene expression changes in response to heat stress, capturing the influence exerted by different organelles upon one nucleus rather than investigating the response of several nuclei in their innate cytoplasmic environments.

In women, breast cancer is the most commonly diagnosed cancer (11.7% of total cases) and the leading cause of cancer death (6.9%) worldwide. Bioactive dietary components such as Sea buckthorn berries are known for their high carotenoid content, which has been shown to possess anti-cancer properties. Considering the limited number of studies investigating the bioactive properties of carotenoids in breast cancer, the aim of this study was to investigate the antiproliferative, antioxidant, and proapoptotic properties of saponified lipophilic Sea buckthorn berries extract (LSBE) in two breast cancer cell lines with different phenotypes: T47D (ER+, PR+, HER2−) and BT-549 (ER-, PR-, HER2−). The antiproliferative effects of LSBE were evaluated by an Alamar Blue assay, the extracellular antioxidant capacity was evaluated through DPPH, ABTS, and FRAP assays, the intracellular antioxidant capacity was evaluated through a DCFDA assay, and the apoptosis rate was assessed by flow cytometry. LSBE inhibited the proliferation of breast cancer cells in a concentration-dependent manner, with a mean IC50 of 16 µM. LSBE has proven to be a good antioxidant both at the intracellular level, due to its ability to significantly decrease the ROS levels in both cell lines (p = 0.0279 for T47D, and p = 0.0188 for BT-549), and at the extracellular level, where the ABTS and DPPH inhibition vried between 3.38–56.8%, respectively 5.68–68.65%, and 35.6 mg/L equivalent ascorbic acid/g LSBE were recorded. Based on the results from the antioxidant assays, LSBE was found to have good antioxidant activity due to its rich carotenoid content. The flow cytometry results revealed that LSBE treatment induced significant alterations in late-stage apoptotic cells represented by 80.29% of T47D cells (p = 0.0119), and 40.6% of BT-549 cells (p = 0.0137). Considering the antiproliferative, antioxidant, and proapoptotic properties of the carotenoids from LSBE on breast cancer cells, further studies should investigate whether these bioactive dietary compounds could be used as nutraceuticals in breast cancer therapy.