Squamous Carcinoma Research Articles

Nomograms for predicting the prognosis in multiple primary esophageal squamous cell carcinoma

Background Due to its rarity, it is challenging to predict the survival of patients with synchronous multiple primary esophageal squamous carcinomas (SMPESCs). We aimed to construct nomograms to predict survival outcomes and help to make therapeutic strategy for patients with SMPESCs. Materials and Methods The clinical and survival data of 135 patients with SMPESCs were analyzed retrospectively. Univariate and multivariate Cox analyses were used to identify independent prognostic factors. Nomograms were constructed to predict 1-year, 3-year and 5-year disease-free survival (DFS) and overall survival (OS). In addition, we further evaluated the effect of postoperative adjuvant therapy on SMPESCs patients with lymph node metastasis. Results In univariate and multivariate analyses of DFS and OS, age, site of the main lesion, lymph node metastasis, total number of lymph nodes dissected, lactate dehydrogenase level and lymphocyte-to-monocyte ratio were identified as independent prognostic factors. These characteristics were further included to establish nomograms. For the internal validation of the nomogram predictions of survival outcomes, the concordance indices were 0.752 and 0.756, respectively. Decision curve analysis also proved the efficacy of the nomograms. Furthermore, adjuvant therapy had a statistically significant benefit for OS but not DFS in patients with lymph node metastasis. Conclusions These nomograms could effectively predict the 1-year, 3-year and 5-year survival outcomes of patients with SMPESCs. Furthermore, adjuvant therapy has the potential to improve OS in patients with lymph node metastasis.

Research progress on the role and mechanism of IGF2BPs family in head and neck squamous carcinoma

Objective:Head and neck squamous cell carcinoma（HNSCC） is one of the common malignant tumours, and most of them are in locally advanced stages at the time of diagnosis due to the lack of early symptoms, and the prognosis of such patients is still poor. M6A modification is the most common form of RNA modification in eukaryotic organisms, with a wide range of biological functions, and the family of IGF2BPs modulates growth, metastasis, chemotherapy resistance, and other processes of cancer by binding to and stabilizing a wide range of target RNAs through recognition of the m6A locus. The aim of this paper is to review the role and related mechanisms of IGF2BPs in head and neck squamous carcinoma, and to provide new ideas for early diagnosis and precision treatment of HNSCC.

Radiology-Pathology Concordance and Prognostication of Nodal Features in pN+ Oral Cavity Cancer.

The aims of our study are to evaluate the diagnostic performance and prognostic value of radiological lymph node (LN) characteristics in pN+ oral cavity squamous carcinoma (OSCC). pN+ OSCC treated between 2012 and 2020 were included. Preoperative imaging was reviewed by a single radiologist blinded to pathologic findings for the following nodal features: imaging-positive LN (iN+), laterality and total number, and image-identified extranodal extension (iENE). The sensitivity of iN+ for pN+ was calculated. The diagnostic performance of other nodal features was evaluated in the iN+ subgroup. The association of radiologic nodal features with overall survival (OS) was evaluated. Inter-rater kappa for radiologic nodal features was assessed in 100 randomly selected cases. Of 406 pN+ OSCC, 288 were iN+. The sensitivity of iN+ for pN+ was 71% overall, and improved to 89% for pN+ LN >1.5 cm. Within iN+, sensitivity/specificity for LN size (>3 cm), total LN number (>4), and ENE were 0.44/0.95, 0.57/0.84, and 0.27/0.96, respectively. Sensitivity of iENE was higher in the subset, with major (>2 mm) versus minor (≤2 mm) pENE (43% vs. 13%, p = 0.001). Reduced OS was observed in iN+ versus iN- (p = 0.006), iENE+ versus iENE- (p = 0.004), LN size >3 versus ≤3 cm (p < 0.001), and higher LN number (p < 0.001). Inter-rater kappa for iN+, laterality, total LN number, and presence of iENE were 0.71, 0.57, 0.78, and 0.69, respectively. Our study shows that despite modest sensitivity of most radiological nodal features, the specificity of image-identified nodal features is high and their prognostic values are retained in pN+ OSCC. 3 (retrospective review comparing cases and controls) Laryngoscope, 134:4947-4955, 2024.

Keratinizing Squamous Cell Carcinoma of Gallbladder Presenting as Hepatic Abscess

Gallbladder tumors are rare but aggressive malignancies, often identified incidentally after cholecystectomy. Adenocarcinoma is the most common phenotype, though squamous cell carcinoma is rare, and it ranged as 1–4% of reported gallbladder cancer. Due to the rarity of the disease, early diagnosis is often challenging. One of the uncommon presentations of gallbladder cancer is hepatic abscess, with few cases reported in the literature. This report presents a rare case of an 82-year-old male referred with hepatic abscess. Multiple pathogens were isolated, indicating a superimposed bacterial infection. Upon further investigation, cytology confirmed the presence of a keratinizing squamous carcinoma of gallbladder cancer. This case presents a rare instance of squamous cell gallbladder carcinoma, which is difficult to diagnose, discovered in an elderly patient who presented with a liver abscess accompanied by bacterial infection. It highlights the challenges in diagnosis and treatment, as well as the poor prognosis associated with the condition.

Lateral forehead flap in oral reconstruction: A case report

Introduction: Lateral Forehead flap is very versatile as it represents a good choice for reparation of different types of nose and oral cavity defects and provides an optimal, aesthetic, and functional outcome. In era of free transfer tissue, the use of loco-regional flaps, alone or in combination may prove to be the only practical solution in the absence of resources or other options. Case presentation: A-65 years old male with multiple comorbidities underwent a wide resection with adequate margin of the lower lip and small segment of the upper lip due to a locally advanced squamous carcinoma. After long consideration, the last resort was the pedicled lateral forehead flap. Information was sourced from patient’s case notes and operating theatre report. Results: The donor defect was covered with a split-thickness skin graft harvested from the right thigh area. The flap survived and both functional and aesthetic outcome were satisfactory. The site of the donor defect result was also acceptable. No complications were noted in our case study beside a small tip necrosis and breakdown of the suture line. Conclusion: Lateral forehead flap is a reliable flap with an acceptable outcome in cases with previous history of radiotherapy or surgery. Decision making of a particular reconstructive option should be done taking in consideration the available resources, surgical team expertise along with the patient's general condition. The pedicled forehead flap still useful when other options are inappropriate or have not been successful.

SMURF1 leads to the β-catenin signaling-mediated progression of esophageal squamous carcinoma by losing PATZ1-induced CCNG2 transcription

Cyclin G2 (CCNG2), a known inhibitor of cell cycle progression, has been identified as a suppressor for the canonical β-catenin pathway. This study explores the impact of CCNG2 on β-catenin activity and malignant characteristics of esophageal squamous cell carcinoma (ESCC) cells, and the mechanism behind CCNG2 dysregulation. In ESCC tissues and cells, CCNG2 was under-expressed and associated with poor clinical outcomes, whereas β-catenin showed an opposite trend. Inducing CCNG2 overexpression in ESCC cells led to a reduction in β-catenin levels, which in turn suppressed proliferation, cell cycle progression, migration, invasion, stemness, and tumorigenesis. Additionally, it enhanced the cytotoxicity and proliferation of T cells in co-culture systems. However, these beneficial effects were negated by the Wnt signaling agonist BML-284. Furthermore, PATZ1 was found as a transcription factor promoting CCNG2 transcription. However, the PATZ1 protein in ESCC cells was degraded by SMURF1. Silencing of SMURF1 restored CCNG2 expression and inhibited β-catenin, thereby suppressing the malignant phenotype of ESCC cells and reducing T cell exhaustion. Yet, these effects were blocked by further silencing of PATZ1. In summary, this research demonstrates that SMURF1 activates β-catenin signaling by suppressing the PATZ1/CCNG2 axis, thereby promoting the progression of ESCC.

Novel Strategies for Lung Cancer Interventional Diagnostics

Lung cancer is a major global health issue, with 2.21 million cases and 1.80 million deaths reported in 2020. It is the leading cause of cancer death worldwide. Most lung cancers have been linked to tobacco use, with changes in cigarette composition over the years contributing to shifts in cancer types and tumor locations within the lungs. Additionally, there is a growing incidence of lung cancer among never-smokers, particularly in East Asia, which is expected to increase the global burden of the disease. The classification of non-small cell lung cancer (NSCLC) into distinct subtypes is crucial for treatment efficacy and patient safety, especially as different subtypes respond differently to chemotherapy. For instance, certain chemotherapeutic agents are more effective for adenocarcinoma than for squamous carcinoma, which has led to the exclusion of squamous carcinoma from treatments like Bevacizumab due to safety concerns. This necessitates accurate histological diagnosis, which requires sufficient tissue samples from biopsies. However, acquiring adequate tissue is challenging due to the complex nature of lung tumors, patient comorbidities, and potential complications from biopsy procedures, such as bleeding, pneumothorax, and the purported risk of local recurrence. The need for improved diagnostic techniques has led to the development of advanced technologies like electromagnetic navigation bronchoscopy (ENB), radial endobronchial ultrasound (rEBUS), and robotic bronchoscopy. ENB and rEBUS have enhanced the accuracy and safety of lung biopsies, particularly for peripheral lesions, but both have limitations, such as the dependency on the presence of a bronchus sign. Robotic bronchoscopy, which builds on ENB, offers greater maneuverability and stability, improving diagnostic yields. Additionally, new imaging adjuncts, such as Cone Beam Computed Tomography (CBCT) and augmented fluoroscopy, further enhance the precision of these procedures by providing real-time, high-resolution imaging. These advancements are crucial as lung cancer is increasingly being detected at earlier stages due to screening programs, which require minimally invasive, accurate diagnostic methods to improve patient outcomes. This review aims to provide a comprehensive overview of the current challenges in lung cancer diagnostics and the innovative technological advancements in this rapidly evolving field, which represents an increasingly exciting career path for aspiring pulmonologists.

Investigating the Cytotoxic Effects of Artemisia absinthium Extract on Oral Carcinoma Cell Line

Background: Artemisia absinthium (A. absinthium), commonly known as absinthe, is a perennial plant with distinctive broad ovate pointed leaves of a silvery-gray color, reaching a height of 1.5 m. The utilization of this herb as a source of natural compounds and as the primary ingredient in the alcoholic beverage absinthe has recently seen a resurgence following a period of prohibition. This study investigates the biological effects of A. absinthium extract on healthy human periodontal ligament stem cells (hPDLSCs) and the human tongue squamous carcinoma cell line (HSC-3). Methods: A. absinthium element characterization was performed using High-Performance Liquid Chromatography (HPLC) and the Folin method. Alizarin assays evaluated the osteogenic capacity of human periodontal ligament cells (hPDLSCs) while CCK-8 and MTT determined the cytotoxicity of the extract against HSC-3 and hPDLSCs. Results: High artemisinin levels were detected, revealing a concentration of 89 μM (25 μg/mL). The total phenolic concentration of the extract was 1.07 mM +/− 0.11. The in vitro cytotoxicity assays revealed the biocompatible profile of the Artemisia extract in hPDLSCs without exhibiting any osteogenic potential. After 24 h of incubation with HSC-3, Artemisia extract (10 µM) decreased cancer cell viability by 99% and artemisinin by 64%, and increased the expression of Caspase 3 and 9 almost six and two times, respectively. Conclusions: In summary, our preliminary findings suggest that A. absinthium extract exhibits a toxic effect against carcinoma cell lines without affecting healthy human periodontal ligament stem cells.

PH-Activatable Molecular Probe for COX-2 Imaging in Human Oral Squamous Carcinoma Cells and Patient-Derived Tissues.

For developing a successful cancer therapeutic modality, the early precise detection of cancer cells in patient biopsies in oral squamous cell carcinoma (OSCC) is crucial. This could help researchers create new diagnostic and therapeutic tools and assist clinicians in recommending more effective treatment plans and improving patient survival. We have developed an SMPD, cyclooxygenase-2 (COX-2) targeting pH-activable fluorophore named CNP, combining a potent COX-2 inhibitor, celecoxib, linked to a naphthalimide fluorophore with an acidic microenvironment-responsive piperazine moiety for specific optical imaging of OSCC in cells and patient tissues. Compared to reference probe RNP lacking celecoxib, CNP selectively enters the COX-2 overexpressing oral cancer cells. Its acidity-responsive imaging response enhances selectivity over cancers with lower COX-2 expression levels and normal cells. Further, CNP is demonstrated in imaging OSCC cells in patient-derived biopsies. Thus, multifunctional CNP shows potential in exploring more reagents for fluorescence-based detection of OSCC cells in patient tissues with translational applications.

Establishing a new human lung squamous cell carcinoma cell line, OMUL‐1, expressing insulin‐like growth factor 1 receptor and programmed cell death ligand 1

AbstractThe Main ProblemSquamous cell carcinoma is the second most prevalent type of non–small cell lung cancer. Analyzing the molecular mechanisms underlying lung carcinoma requires useful tools, such as squamous lung cancer cell lines.MethodsA novel new lung squamous cell carcinoma cell line, OMUL‐1, was developed from the primary lung cancer of a 74‐year‐old man. We assessed the characteristics and behavior of OMUL‐1 cells were examined, including their growth kinetics, tumorigenicity in mice, histological properties, gene expression profiles using reverse transcription polymerase chain reaction (RT‐PCR), and RNA sequencing and invasion assays.ResultsOMUL‐1‐an adherent cell line‐resulted in 100% tumor formation when subcutaneously injected into mice. Histological analysis of the subcutaneous tumor using hematoxylin and eosin staining revealed squamous cell carcinoma with characteristics similar to those of the primary tumor (p40 and p63 were positive, and TTF‐1 was negative). An invasion assay demonstrated that OMUL‐1 had a lower invasion ability compared to that of other developed cell lines. RT‐PCR analysis and RNA sequencing indicated that OMUL‐1 cells expressed FGFR1, FGFR2, FGFR3, FGFR4, EGFR, HER2, ErbB3, ErbB4, VEGFR3, IGF1R, c‐MET, PDGFRa, and PDGFRb. Additionally, picropodophyllin (an IGF1R inhibitor) significantly inhibited the growth of OMUL‐1 cells. Immunohistochemistry revealed that IGF1R and PD‐L1 were expressed in both the primary and subcutaneous tumors.ConclusionsWe developed a novel new squamous cell lung carcinoma cell line, OMUL‐1, that expresses IGF1R and PD‐L1.

Abstract C010: Tumor MK2 drives tumor progression and an immunosuppressive microenvironment in head and neck squamous cell carcinoma

Abstract Background: Head and neck squamous carcinoma (HNSCC) results in poor patient outcomes driven primarily by locoregional and distant metastatic tumor spread. We have demonstrated that highly phosphorylated/active MAPKAPK-2 (MK2) is a poor prognostic factor in HNSCC patients. MK2 is associated with the expression of pro-inflammatory cytokines that are linked with creating a pro-tumorigenic microenvironment. Nevertheless, how MK2 contributes to HNSCC development has not been established. In this study, we aimed to identify the effect of MK2 on cancer progression, immune infiltration, and cytokine secretion. Method: To assess the effect of tumor MK2 in vivo, we used CRISPR-Cas9 to knock out (KO) MK2 in a metastatic murine HNSCC cell line, Ly2, and orthotopically implanted wildtype (WT) or MK2 KO cells into the floor-of-mouth of Balb/c mice. At the endpoint of the experiment, the tumors were excised, and myeloid and lymphocyte immune profiling was done via flow cytometry. Mouse lungs and cervical lymph nodes were dissected, paraffin-fixed, sectioned, and H&amp;E stained, and the presence/quantity of metastatic foci was evaluated by a blinded board-certified pathologist. We broadly evaluated the tumor secretome via a 105-cytokine dot blot array using 8 mg of the same resected tumor sample and performed densitometric analysis comparing WT and KO tumors. Results: Loss of MK2 significantly reduced primary tumor growth, as well as lymph node and lung metastasis, compared to WT tumors. Tumor immunophenotyping by flow cytometry revealed that MK2 KO tumors had increased overall immune cell (CD45+) infiltration, but decreased quantities of immunosuppressive neutrophils (CD11b+Ly6G+Arg1+Ly6C-) and a shift in macrophage polarity favoring M1 (anti-tumor) macrophages (CD11b+Ly6G-F4-80+CD80+). In addition, there were increased quantities of anti-tumor immune cells including natural killer (NK)-T, CD4+, and CD8+ T cells in the MK2-KO tumors, while the proportion of CD4+CD25+ Tregs and exhausted CD8 T cells (CD8+PD1+) was decreased compared to WT tumors. Furthermore, cytokine array analysis revealed that secreted levels of several cytokines, including CXCL1, CXCL2, GM-CSF, G-CSF, IL-1alpha, and IL-6, were significantly lower in MK2 KO tumors compared to WT tumors. Conclusion: These results show that tumoral MK2 signaling promotes tumor growth, metastasis, immunosuppressive inflammation, and pro-tumorigenic cytokine release which substantiates future therapeutic targeting of this pathway. Citation Format: Dakota D.D. Okwuone, Deri Morgan, Alyssa Schmidt, Devin Shrock, Yuting Lin, Hao Gao, Sufi M. Thomas, Gregory N. Gan. Tumor MK2 drives tumor progression and an immunosuppressive microenvironment in head and neck squamous cell carcinoma [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Tumor-body Interactions: The Roles of Micro- and Macroenvironment in Cancer; 2024 Nov 17-20; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2024;84(22_Suppl):Abstract nr C010.

Abstract PR018: Inflammation-induced mechanotransduction is necessary and sufficient to create pre-cancerous squamous lung metaplasias and necessary to drive progression to dysplasia

Abstract Chronic exposure to tobacco smoke triggers chronic inflammation, fostering a fibrotic microenvironment and promoting the focal replacement of columnar human bronchial epithelial cells (hBECs) with cells exhibiting squamous morphology, termed metaplasia. How squamous metaplasia progresses to squamous dysplasia and, ultimately, squamous lung carcinoma is under intense investigation. Using human bronchial tissues, we demonstrate that, both in vitro and in vivo, cellular stress associated with chronic inflammation (e.g., oxidative stress, DNA damage) enhances TGF-β signaling and creates a fibroblast state exhibiting upregulated HSP47 expression, driving increased collagen fiber alignment, elevated tissue stiffness, and subsequent activation of YAP-dependent mechanotransduction in adjacent hBECs. These Stress/Tension-producing Instructive Fibroblasts (STIFs) alone were sufficient to induce hBECs to squamous metaplasia, precursors to tumor formation. Furthermore, when tumor suppressor function was compromised in hBECs, STIFs induced dysplastic phenotypes. Mechanistically, STIFs act in a dominant manner modulating epithelial cell identity and malignant phenotypes via extracellular matrix (ECM)-dependent mechanotransduction. Inhibition of HSP47-dependent collagen processing effectively prevented fibroblast-induced squamous metaplasia and dysplasia and was capable of reversing fibroblast-induced metaplasia, restoring bronchial epithelial cell identity. Citation Format: Thea D. Tisty, Deng Pan, Philippe Gascard, Joseph Caruso, Lorenzo Ferri, Chira Chen-Tanyolac. Inflammation-induced mechanotransduction is necessary and sufficient to create pre-cancerous squamous lung metaplasias and necessary to drive progression to dysplasia [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Tumor-body Interactions: The Roles of Micro- and Macroenvironment in Cancer; 2024 Nov 17-20; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2024;84(22_Suppl):Abstract nr PR018.

Abstract B015: Targeting CX3CL1 transforms a cold tumor microenvironment into a hot one by enhancing T cells and DCs locally and systemically

Abstract Intratumoral heterogeneity (ITH) is associated with immunotherapy failure and a weaker anti- tumor immune response. Using a series of carcinogen-induced tumor (CIT) squamous cell skin carcinoma cell lines, our lab has sought to identify tumor-intrinsic factors—particularly chemokines and cytokines—which can have a “dominant negative” effect on the immune microenvironment when present within a heterogeneous tumor. We identified CX3CL1 as a one such critical mediator of a cold (T cell low) immune microenvironment and found that tumors containing as few as 10% CX3CL1high tumor cells exhibit a diminished T cell response compared to tumors which lack CX3CL1high tumor cells. We predicted that targeting CX3CL1 could therefore transform a cold tumor microenvironment (TME) into an inflammatory hot (T cell high) TME. Supporting this, we found that CRISPR-Cas9-mediated deletion of CX3CL1 in a tumor cell line that typically gives rise to cold tumors led to higher T cell infiltration and fewer immunosuppressive macrophages in the TME of CX3CL1-/- tumors. Building on this, we further explored T cell populations in the TME and found that CX3CL1-/- tumors had significantly increased activation and decreased exhaustion markers on T cells. Flow cytometry analysis showed higher expression of CD44 on effector CD4 T cells and an increase in the number of activated CD44+ PD1low CD4 T cells. By contrast, the frequency of CD4 T cells which were PD1High TIM3+ and PD1High TIM3+ TIGIT+ which represent a likely exhausted effector CD4 population was lower in CX3CL1-/- tumors than in counterpart control tumors. An interrogation of DC subsets in the TME further revealed a higher number of cDC1 and cDC2 populations in CX3CL1−/− tumors. To determine the effect of tumor-produced CX3CL1 on immune activity in lymphoid organs, we investigated T cell and DC markers in the spleen and tumor-draining lymph nodes during early tumor development. Spleens harvested 10 days after tumor injection showed higher infiltration of cDC1 CCR7+ and cDC2 CCR7+ mature DCs in CX3CL1-/- tumor-bearing mice. Thus, in the absence of tumor-produced CX3CL1, both cDC1 and cDC2 dendritic cell subsets attain more mature phenotypes in both the TME and the spleen. At the same time point, tumor-draining lymph nodes also exhibited a shift in immune cell composition characterized by an increased frequency of T cells, particularly CD4+ T cells, and a decreased frequency of B cells. Moreover, the percentage of PD1+ CD8 cells was also higher, indicating better priming and activation of CD8 T cells via DCs in tumor-draining lymph nodes. These findings indicate that tumor-produced CX3CL1 drives a cold immune microenvironment through both tumor-localized and systemic effects. These studies suggest therapeutically targeting CX3CL1 particularly in tumors where tumor cells produce high levels of CX3CL1 holds promise to transform a cold TME into a hot immune inflammatory TME and potentiate better anti-tumor immunity. Citation Format: Piyush Chaudhary, Savannah Hughes, Joshua Tay, Robert Letchworth, Miho Tanaka, Melissa Reeves. Targeting CX3CL1 transforms a cold tumor microenvironment into a hot one by enhancing T cells and DCs locally and systemically [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Tumor-body Interactions: The Roles of Micro- and Macroenvironment in Cancer; 2024 Nov 17-20; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2024;84(22_Suppl):Abstract nr B015.

A Six-Year Examination of the Influence of Surgical Technique and Intraoperative Intraglandular Clostridium Botulinum Toxin Application in Salivary Gland Tumor Operations.

Introduction: Salivary gland tumor operations are associated with complications including facial nerve dysfunction (FND) and salivary fistula. The objective of this study was to investigate the effect of extracapsular dissection (ECD) and the application of Clostridium botulinum toxin (CBT) in contrast to partial and lateral parotidectomy on complications. Methods: All salivary gland tumor operations performed within the last 6 years were retrospectively examined. Data were collected from electronic patient files from our otorhinolaryngology clinic. Total parotidectomies and submandibulectomies were not included in the analysis of CBT application. Results: In total, 418 cases were examined, including 84 (20%) malignant tumors. In total, 18 patients underwent ECD, 93 partial parotidectomy, 199 lateral parotidectomy, 76 total parotidectomy, and 32 submandibulectomy. The most common complication was transient FND (49%; n = 205; data available for 415 patients), which was measured at four days. Additional complications included salivary fistula (n = 56), infection (n = 49), bleeding or hematoma (n = 21). Preoperative facial nerve paralysis (p < 0.0001), pain (p < 0.0001), and a history of squamous cell skin carcinoma (SCC) (p < 0.001) were predictive of malignancy. The application of CBT did not reduce the risk of salivary fistula (p-value: 0.0182) and was associated with a higher combined complication rate (p-value: 0.0199). ECD was not associated with a lower likelihood for FND (p = 0.350). Conclusions: Preoperative pain, facial paralysis, or a history of SCC are predictors of malignancy. Use of CBT was not associated with a reduced risk of salivary fistula, but rather a higher complication rate.

Effect of hinokitiol in ameliorating oral cancer: in vitro and in silico evidences.

The study aimed to evaluate the anticancer potential of hinokitiol in treating oral cancer by using in vitro models and examining its interaction with the Pim-1 protein through in silico methods. Hinokitiol was applied to KB-1 oral squamous carcinoma cells, where the half-maximal inhibitory concentrations (IC50) was determined. Morphologic changes in treated cells were observed using phase contrast microscopy, while acridine orange/ethidium bromide (AO/EB) staining was used to assess nuclear changes and apoptosis. Flow cytometry was employed to analyze the cell-cycle progression. Given the high expression of Pim-1 in oral squamous carcinoma cells, molecular docking and simulation were performed to evaluate hinokitiol's binding affinity and stability with the Pim-1 protein. To compare its effects, hinokitiol was also tested on non-cancerous pre-adipocytes (3T3-L1), providing insights into its selective cytotoxicity between healthy and cancerous cells. Hinokitiol treatment resulted in cytotoxic effects on KB-1 oral squamous carcinoma cells, with an IC50 of 30µg/mL after 24 and 48hs of exposure. Morphologic studies showed reduced cell population and density. In contrast, hinokitiol exhibited lower toxicity and caused fewer morphological changes in non-cancerous 3T3-L1 pre-adipocytes. Apoptosis was confirmed through acridine orange/ethidium bromide staining, while flow cytometry revealed cell-cycle arrest in the Synthesis phase (S) and Gap 2 phase/ Mitosis Phase (G2/M) phases. Molecular docking showed strong binding of hinokitiol to Pim-1, and simulations confirmed the interaction's stability. These findings suggest hinokitiol selectively targets cancer cells and effectively inhibit Pim-1, supporting its potential as an oral cancer treatment.

OGC SO03 - Impact of recent European consensus guidelines on Oligometastic Disease (OMD) and potential case load of Oesophagogastric cancer resections

Abstract Background To evaluate the impact of recently devised oligometastatic disease (OMD) criteria as per the European consensus guidelines on the eligibility of Oesophagogastric (OG) cancer patients for curative treatment. Method A retrospective analysis was conducted on OG cancer cases discussed at multidisciplinary team (MDT) meetings at a tertiary university hospital from January 2018 to December 2023. Inclusion criteria included T2-T4a adenocarcinomas, T2-T4b squamous cell carcinomas, small cell carcinoma and adeno-squamous carcinomas that were classified as M1 disease. Exclusion criteria were T4b adenocarcinomas, M1 peritoneal disease, non-adeno/ squamous carcinoma histology, M1 peritoneal cytology, and head and neck squamous cell carcinomas proximal to the thoracic inlet. Data on patient demographics, T stage, histology, and metastatic sites were analysed to identify those meeting the oligometastatic disease criteria. Results 483 of 696 were diagnosed as M1 disease. OMD was found in 59 cases (12.2%), with a median age of 72 years (43 males, 16 females). T stage distribution: 3 T2, 36 T3, 20 T4. Histology: 39 adenocarcinomas, 19 squamous cell carcinomas, 1 small cell carcinoma. Metastatic sites: non-regional lymph nodes (77.9%), liver (13.6%), lung (5.1%), bone (1.7%), adrenal (1.7%) Location of Lymph Node Involvement: 1Para-aortic11 (23.9%)2Right supra-clavicular7 (15.2%)3Porto-caval6 (13.0%)4Retro-crural5 (10.9%)5Upper mediastinal5 (10.9%)6Left Supraclavicular5 (10.9%)7Coeliac4 (8.6%)8SMA3 (6.6%) Conclusion Our data would suggest that 12.2% of patients discussed in the MDT may potentially be candidates for a curative intent treatment pathway. Although the long-term outcomes remain unclear, this approach may define a subgroup of patients who may benefit from curative intent treatment with the highest site being para-aortic nodes.

GPD1L may inhibit the development of esophageal squamous cell carcinoma through the PI3K/AKT signaling pathway: bioinformatics analysis and experimental exploration.

Esophageal squamous carcinoma (ESCC) is the most prevalent pathological subtype of esophageal cancer (EC). It has the characteristics of significant local invasion, quick disease progression, high recurrence rates, and a dismal prognosis for survival. Phosphatidylinositol 3-kinase/serine-threonine kinase (PI3K/AKT) is a signaling system whose aberrant activation regulates downstream factors, leading to the promotion of cancer development. This study looks at a protein called Glycerol-3-phosphate dehydrogenase 1-like (GPD1L), which strongly affects the development of several cancers. However, its association with ESCC development and its underlying mechanisms are not clear. In this paper, we analyzed six ESCC transcriptome data obtained from the GEO database. We utilized bioinformatics technology and immunohistochemistry to differentially analyze GPD1L levels of mRNA and protein expression in ESCC and normal adjacent tissues. Furthermore, we conducted survival, co-expression, enrichment, immune infiltration and drug sensitivity analysis. Moreover, we further investigated the role and mechanism of GPD1L by Western Blot (WB), Cell Counting Kit-8 (CCK8), wound healing assay, Transwell assay, and flow cytometry. Finally, the addition of IGF-1, the activator of PI3K/AKT, could rescue the inhibitory effect of GPD1L on ESCC. The findings manifest that the expression of GPD1L was low in ESCC, and functional experiments showed that GPD1L promoted apoptosis in vitro while blocking cell migration, invasion, and proliferation. Based on mechanism research, GPD1L's impact on ESCC could be explained by its suppression of the PI3K/AKT signaling pathway's activation. To sum up, our findings imply that GPD1L may impede the initiation and advancement of ESCC via modulating the PI3K/AKT signaling pathway. GPD1L is considered to be a promising therapeutic target and biomarker to diagnose and treat ESCC.

Forkhead box N1 is possibly a novel biomarker and prognostic indicator for patients with squamous cell lung carcinoma.

Background: The roles of Forkhead box N1 (FOXN1) in lung squamous cell carcinoma (LUSC) remains elusive. This study was focused on assessing the expression levels of FOXN1 in LUSC and exploring its potential clinical implications. Methods: Utilizing a range of databases, this study conducted an analysis of the FOXN1 gene's expression levels, comparing LUSC samples with those from normal lung tissues. The expression levels of FOXN1 in primary LUSC and corresponding normal lung tissues were assessed using immunohistochemistry (IHC). Histoscore was used to evaluate the staining degree. χ2 test and Fisher's exact test were employed to assess the association between categorical variables that do not possess an ordinal nature. Multivariate survival analysis was conducted using the Kaplan-Meier method, the Wilcoxon test, and the Cox proportional hazards model. Results: In contrast to normal lung tissues, the expression of the FOXN1 gene was found to be significantly elevated in LUSC tissues (P < 0.01). And FOXN1 was expressed in 79 (98.8%) evaluated LUSC tissues, most of which showed compositive IHC-staining intensity, presenting heterogeneously expression. 69 (87.3%) cases were characterized for strong immunostaining intensity, 70 (87.5%) cases showed moderate intensity, and 66 (82.5%) cases presented weak intensity. Only one sample of normal lung tissue, which represents 10% of the total, exhibited weak immunostaining exclusively (P < 0.05). Additionally, the expression of FOXN1 was found to have a significant correlation with the grading of LUSC, the presence of lymph node and distant metastases, the stage of the disease, and the survival outcomes (P < 0.05). Conclusion: The expression of FOXN1 is frequently increased in LUSC, and the patients with high FOXN1 expression have a poorer survival outcome. FOXN1 can be a novel biomarker and prognostic indicator for LUSC patients.

Effects of Pseudomonas aeruginosa pyocyanin and 1-hydroxyphenazine on intracellular calcium, mitochondrial function, and viability in human nasal epithelial cells.

Pseudomonas aeruginosa is an opportunistic pathogen that produces phenazine metabolites pyocyanin and 1-hydroxyphenazine that have been suggested to have detrimental effects on mitochondrial function and reactive oxygen species (ROS) production. Prior studies have suggested activation of Ca 2+ signaling by pyocyanin in an airway cell line. Ca 2+ is tightly linked to both normal mitochondrial function as well as mitochondrial ROS and apoptosis during mitochondrial Ca 2+ overload. We found that pyocyanin but not 1-hydroxyphenazine induced both cytosolic and mitochondrial Ca 2+ increases. Despite this, both pyocyanin and 1-hydroxyphenazine decreased in cell viability in RPMI 2650 nasal carcinoma cells but not in primary human nasal epithelial cells (HNECs) at 24 hours. However, in both RPMI 2650 and HNEC, mitochondrial membrane potential acutely decreased after treatment with either pyocyanin or 1-hydroxyphenazine. Our data suggest that Ca 2+ signaling is not required for acute effects of 1-hydroxyphenazine or pyocyanin on mitochondrial function. The greater sensitivity of RPMI 2650 cells to pyocyanin-induced and 1-hydroxyphenzine-induced cytotoxicty compared with primary cells suggests that these compounds might have some applicability in treating nasal squamous carcinoma or other types of head and neck squamous carcinoma.

Jing-Si herbal tea attenuates oral cancer cell viability, migration, and invasion by regulating epithelial-to-mesenchymal transition and reactive oxygen species accumulation

IntroductionOral cancer is the sixth most common cancer worldwide and can be life-threatening if not diagnosed and treated in its early stages. Jing-Si herbal tea (JSHT), containing eight traditional Chinese medicine-based herbs, is known to suppress the malignancy, growth, and metastasis of several tumour cells, including breast, lung, and colon cancer cells. However, the pharmacological effect of JSHT on oral cancer progression is still unclear. MethodsIn this study, we evaluated the potential of JSHT to arrest the development of oral cancer via induction of cell death by reactive oxygen species (ROS) accumulation and inhibition of migration, invasion, and epithelial-to-mesenchymal transition (EMT). For this purpose, we employed the human tongue squamous cell carcinoma human tongue squamous carcinoma cell line, which we treated with different JSHT concentrations for 24 hours, and assessed cell viability, migration, and wound healing capabilities, together with western blotting for measuring expression levels of EMT markers. ResultsSurvival, migration/invasion ability, EMT, and ROS production in the human tongue squamous carcinoma human tongue squamous carcinoma and FaDu human pharynx squamous cell carcinoma cell line were decreased by JSHT treatment via Lon protease-independent mechanisms. ConclusionThis study demonstrates that JSHT could be regarded as a candidate new supplement to existing anticancer therapies and an alternative, orally administered healthcare product for treating oral squamous cell carcinoma.