Case report: EGFR fusion mutation combined with EGFR amplification responds to EGFR-TKI therapy.

Given their good antitumor effects, epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKIs) are standard first-line therapy for EGFR-sensitive mutations, including exon 19 deletions and exon 21 L858R mutations. EGFR fusion mutations and EGFR amplification are very rare in non-small cell lung cancer (NSCLC). We describe 2 patients with NSCLC harboring EGFR fusion mutations (EGFR-MACF1 and EGFR-GNAT3) combined with EGFR amplification. Both patients received EGFR-TKI treatment, and 1 of them showed an antitumor response.

FOXA1/UBE2T Inhibits CD8+T Cell Activity by Inducing Mediates Glycolysis in Lung Adenocarcinoma.

BACKGROUND: Immune escape is a key factor influencing survival rate of lung adenocarcinoma (LUAD) patients, but molecular mechanism of ubiquitin binding enzyme E2T (UBE2T) affecting immune escape of LUAD remains unclear. The objective was to probe role of UBE2T in LUAD. METHODS: Bioinformatics means were adopted for analyzing UBE2T and forkhead box A1 (FOXA1) expression in LUAD tissues, the gene binding sites, the pathway UBE2T regulates, and the correlation between UBE2T and glycolysis genes. Dual luciferase and chromatin immunoprecipitation (ChIP) assays were conducted for validating the binding relationship between the two genes. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) and western blot were employed to evaluate UBE2T, FOXA1, and programmed death ligand 1 (PD-L1) levels in cancer cells. MTT assay was conducted for detecting cell viability. Cytotoxicity assay detected CD8+T cell toxicity. Cytokine expression was assayed by enzyme linked immunosorbent assay (ELISA). Extracellular acidification rate (ECAR) and oxygen consumption rate (OCR) were assayed by extracellular flow analyzer. Glycolytic gene expression was analyzed by qRT-PCR, and glycolysis-related indicators were detected by ELISA. Immunohistochemistry (IHC) detected CD8+T cell infiltration in tumor tissues. RESULTS: FOXA1 and UBE2T were up-regulated in LUAD, and a binding site existed between UBE2T and FOXA1. Overexpressing UBE2T could increase PD-L1 expression and inhibit toxicity of CD8+T cells to LUAD cells. Overexpressing UBE2T repressed CD8+T cell activity in LUAD by activating the glycolysis pathway, and the addition of glycolysis inhibitor 2-deoxy-d-glucose (2-DG) reversed the above results. Mechanistically, FOXA1 promoted the immune escape of LUAD by up-regulating UBE2T and thus mediating glycolysis. In vivo experiments revealed that UBE2T knockdown hindered tumor growth, inhibited PD-L1 expression, and facilitated CD8+T cell infiltration. CONCLUSION: FOXA1 up-regulated the expression of UBE2T, which activated glycolysis, and thus inhibited activity of CD8+T cells, causing immune escape of LUAD.

Aumolertinib: effective treatment for asymptomatic pulmonary giant cell carcinoma with EGFR L858R mutation - a case report.

Aumolertinib, as a novel third-generation epidermal growth factor receptor tyrosine kinase inhibitor (EGFR-TKI), has been widely employed as a first-line treatment for advanced non-small cell lung cancer (NSCLC) patients with EGFR mutation. However, reports regarding the benefit of using aumolertinib as a monotherapy in pulmonary giant cell carcinoma are relatively scarce. In this report, we present a pulmonary giant cell carcinoma case harboring the EGFR Leu858Arg (L858R) mutation, with the patient at stage cT2bN3M1c IVB. Through the use of autolearning as a single agent, we effectively controlled the progression of pulmonary giant cell carcinoma, achieving a 6-month progression-free survival during the treatment course. Notably, the patient's tumor not only ceased its growth but also continued to shrink, highlighting a significant therapeutic effect. This case reveals the effectiveness of aumolertinib as a monotherapy in controlling disease progression. The finding underscores the therapeutic advantage of aumolertinib in this particular subgroup of patients, offering a novel treatment option for pulmonary giant cell carcinoma.

Construction of a prognostic model for lung adenocarcinoma based on membrane-tension-related genes.

Background: Lung adenocarcinoma (LUAD), which is the most common type of non-small cell lung cancer (NSCLC), is one of the most aggressive and fatal tumors. Therefore, the identification of key biomarkers affecting prognosis is important to improving the prognosis of patients with LUAD. Cell membranes have long been understood; however, few studies have focused on the role of membrane tension in LUAD. The present study aimed to construct a prognostic model associated with membrane-tension-related genes (MRGs) and explore its prognostic value in LUAD patients. Methods: RNA sequencing data and the corresponding clinical characteristics data of LUAD were obtained from The Cancer Genome Atlas (TCGA) database. Five membrane-tension prognosis-related genes (5-MRG) were screened by univariate and multifactorial COX regression and least absolute shrinkage and selection operator (LASSO) regression analyses. The data were then divided into testing, training, and all groups to build a prognostic model, and Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), copy number variations (CNV), tumor mutation burden (TMB), and tumor microenvironment (TME) analyses were performed to explore the potential mechanisms of MRGs. Finally, single-cell data from the GSE200972 dataset in the Gene Expression Omnibus (GEO) database were obtained to determine the distribution of prognostic MRGs. Results: Construction and validation of the prognostic risk models were conducted using 5-MRG in the trial, test, and all data sets. Patients in the low-risk group had a better prognosis than those in the high-risk group, and the Kaplan-Meier survival curve and receiver operating characteristic curve (ROC) confirmed that the model had a better predictive value for LUAD patients. GO and KEGG analyses of differential genes in the high- and low-risk groups were significantly enriched in immune-related pathways. Immune checkpoint (ICP) differential genes differed significantly in the high- and low-risk groups. By analyzing the single-cell sequencing data, the cells were divided into nine subpopulations and cell subpopulation localization through 5-MRG. Conclusions: The results of this study suggest that a prognostic model based on prognosis-associated MRGs can be used to predict the prognosis of LUAD patients. Therefore, prognosis-related MRGs could be potential prognostic biomarkers and therapeutic targets.

Expression of Polyadenylate-binding Protein Cytoplasmic 1 (PABPC1) in Combination With RAD51 as Prognostic Biomarker in Patients Who Underwent Postoperative Chemotherapy for Esophageal Squamous Cell Carcinoma.

Molecular markers in the prognosis of esophageal squamous cell carcinoma (ESCC) patients who received postoperative treatments are lacking. This research aims to evaluate the prognostic value of polyadenylate-binding protein cytoplasmic 1 (PABPC1) alone and in combination with RAD51 in ESCC patients who underwent postoperative chemotherapy (CT). A total of 103 ESCC patients who underwent postoperative CT and 103 matched ones who received surgery alone were analyzed in this study. PABPC1 and RAD51 expression was assessed in cancer samples by immunohistochemistry. PABPC1 high expression (PABPC1-HE) but not that of RAD51 was associated with poor patients' survival, regardless of the postoperative treatment or node status. Patients with PABPC1 low expression and RAD51 negative expression [RAD51- (PABPC1-LE/RAD51-)] tumor had good overall survival (OS) in both the CT treated and untreated groups. Patients with PABPC1-LE/RAD51+ and PABPC1-HE/RAD51+ tumors had longer OS in the CT treated group than in the untreated group. However, PABPC1-HE/RAD51- was associated with a poor outcome in both groups and the patients with PABPC1-HE/RAD51- tumor had hardly any benefit from CT in N+ status. PABPC1 alone and in combination with RAD51 was a prognostic biomarker for OS in ESCC patients who received postoperative CT.

Construction of a prognostic model for lung squamous cell carcinoma based on immune-related genes.

Lung squamous cell carcinoma (LUSC) lacks appropriate prognostic and diagnostic strategies. Available studies suggest the effectiveness of immunotherapy for LUSC, but effective molecular markers are still insufficient. We obtained mRNA expression and clinical information of LUSC samples from The Cancer Genome Atlas (TCGA) database. Enrichment levels of immune-related genes were revealed by single sample gene set enrichment analysis. Then, differentially expressed genes (DEGs) related to immunity were obtained by differential analysis. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were performed. In addition, Cox regression analysis combined with LASSO method was utilized to identify immune-related prognostic genes, and an immune-related prognostic model was constructed. Kaplan-Meier and receiver operating characteristic (ROC) curves were drawn to verify the accuracy of the model. Finally, a nomogram and calibration curve were drawn to predict LUSC patients' survival. Samples were assigned into high-, medium- and low-immune groups. Compared with low- and medium-immune groups, high-immune group enriched more immune cells, with higher immune infiltration degree, and higher expression of immune checkpoints and human leukocyte antigen. DEGs were enriched in biological processes and signaling pathways related to immunity. Eleven genes (ONECUT3, MAGED4, SULT2A1, HPR, S100A5, IRS4, DPP6, FGF8, TEX38, PLAAT1 and CLEC3A) were obtained to construct an immune-related prognostic model. Riskscore served as an independent prognostic factor. Besides, the nomogram prediction model could predict disease progression in LUSC patients. The constructed risk assessment model for LUSC immune-related genes could assess LUSC patients' prognoses with great efficacy, providing guidance for the clinical treatment of LUSC.

Systematic analysis of apoptosis-related genes in the prognosis of lung squamous cell carcinoma: a combined single-cell RNA sequencing study.

Background: Lung squamous cell carcinoma (LUSC) has a poor prognosis and lacks appropriate diagnostic and treatment strategies. Apoptosis dysregulation is associated with tumor occurrence and drug resistance, but the prognostic value of apoptosis-related genes (ARGs) in LUSC remains unclear. Methods: Using univariate Cox regression, least absolute shrinkage and selection operator (LASSO) regression, and multivariate Cox regression analysis based on differentially expressed ARGs, we constructed an ARG-related prognostic model for LUSC survival rates. We conducted correlation analysis of prognostic ARGs by incorporating the dataset of normal lung tissue from the Genotype-Tissue Expression (GTEx) database. We then constructed a risk model, and the predictive ability of the model was evaluated using receiver operating characteristic (ROC) curve analysis. Non-small cell lung cancer (NSCLC) single-cell RNA sequencing (scRNA-seq) data were downloaded from the Gene Expression Omnibus (GEO) database. Subsequently, these data were subjected to single-cell analysis. Cell subgroups were determined and annotated by dimensionality reduction clustering, and the cell subgroups in disease development were identified via pseudotemporal analysis with the Monocle 2 algorithm. Results: We identified four significantly prognostic ARGs and constructed a stable prognostic risk model. Kaplan-Meier curve analysis showed that the high-risk group had a poorer prognosis (P<0.05). Furthermore, the ROC analysis of 3-, 5- and 7-year survival rates confirmed that the model had good predictive value for patients with LUSC. Single-cell RNA sequencing showed the prognostic ARGS were enriched in epithelial cells, smooth muscle cells, and T cells. Pseudotime analysis was used to infer the differentiation process and time sequence of cells. Conclusions: This study identified ARGs that are associated with prognosis in LUSC, and a risk model based on these prognostic genes was constructed that could accurately predict the prognosis of LUSC. Single-cell sequencing analysis provided new insights into the cellular-level development of tumors. These findings provide more guidance for the diagnosis and treatment of patients with LUSC.

Transient receptor potential vanilloid 4 promotes the growth of non-small cell lung cancer by regulating Foxp3.

OBJECTIVE(S): Transient receptor potential vanilloid 4 (TRPV4) participates in malignant tumor. However, the role of TRPV4 in non-small cell lung cancer (NSCLC) remains unclear. In this study, we demonstrated TRPV4 was upregulated in NSCLC tissues and NSCLC cell lines. MATERIALS AND METHODS: TRPV4 level in the NSCLC patients and cell lines were detected, and its function was studied both in vivo and vitro. RESULTS: The level of TRPV4 showed a positive correlation with tumor size of NSCLC patients. Activation TRPV4 by agonist GSK1016790A promoted cell proliferation and decreased apoptosis in A549 cells, and these effects were enhanced when the cells have overexpressed TRPV4. Moreover, GSK1016790A induced inhibitory effects on apoptosis of A549 cells was impaired when GSK1016790A used together with TRPV4 selective antagonist HC-067047, or impaired when the cells have already downregulated TRPV4 expression by TRPV4 siRNA. In vivo study, pharmacological inhibition of TRPV4 prevented A549 cells transplanted tumor growth. It was showed Foxp3 level was significantly increased in the NSCLC tissues, and showed a positive correlation with the level of TRPV4. Deactivation of TRPV4 using TRPV4 siRNA or HC-067047 significantly reduced expression of Foxp3 in GSK1016790A treated NSCLC cells. Moreover, downregulation Foxp3 by transfection of Foxp3 siRNA significantly impaired TRPV4 induced NSCLC cells proliferations in vitro. CONCLUSIONS: Antitumor effects caused by TRPV4 inhibition in NSCLC might be attributed to the suppression of Foxp3 which induced subsequent cell apoptosis. Thus, pharmacological inhibition of TRPV4 may be a promising option for NSCLC treatment.

High-Expression of Cytoplasmic Poly (A) Binding Protein 1 (PABPC1) as a Prognostic Biomarker for Early-Stage Esophageal Squamous Cell Carcinoma.

BACKGROUND AND OBJECTIVE: Poly (A) binding protein cytoplasmic 1 (PABPC1) plays a crucial role in the regulation of RNA polyadenylation, translation initiation, and mRNA stability and may be involved in tumorigenesis. Herein, we set out to identify the prognostic value of PABPC1 expression in esophageal squamous cell carcinoma (ESCC). METHODS: Using quantitative real-time PCR (qRT-PCR) and immunohistochemical analysis, the present study investigated mRNA and protein expressions of PABPC1 in 231 ESCCs and their paired adjacent normal epithelial tissues. RESULTS: We observed a reduction in the average mRNA expression of PABPC1 in ESCC tissue specimen, but the mRNA expression of PABPC1 was significantly higher (P<0.001) in ESCC tissues with high PABPC1 expression and lower (P=0.033) in tissues with low PABPC1 expression. In immunohistochemical analysis, positive expression of the PABPC1 protein was identified in 179 ESCC tissue specimens (179/231, 77.5%), while the percentage of ESCC tissue specimens with high expression of PABPC1 was found to be 41.1% (95/231). PABPC1 expression was found to be significantly correlated with lymph node metastasis (LNM) (P=0.011), pathological stage (P=0.021), tumor recurrence (P<0.001), and the outcome (P<0.001) of patients with ESCC. High expression of PABPC1 was associated with poor overall survival (OS) of ESCC patients (P<0.001) among all pathological stages, particularly in the early stages (pStage-I and -II), and identified to be an independent prognostic factor for OS of patients with ESCC in multivariate analysis (HR=2.622; 95% CI, 1.68-4.129). Comparatively, the expression of Ki-67, p53, and nm23 was not associated with OS. CONCLUSION: In this study, we discovered that PABPC1 is a prognostic biomarker and a therapeutic target for ESCC, particularly early-stage ESCC.

MiR-654-3p Suppresses Non-Small Cell Lung Cancer Tumourigenesis by Inhibiting PLK4.

PURPOSE: MiR-654-3p plays important roles in many types of malignant tumours. However, the biological function of miR-654-3p in non-small cell lung cancer (NSCLC) remains unknown. In this study, the role of miR-654-3p in NSCLC was investigated. METHODS: qRT-PCR was used to evaluate the level of miR-654-3p in NSCLC tissues and cell lines, while Cell Counting Kit-8, Annexin V/propidium iodide dual staining or TUNEL staining were used to investigate proliferation and apoptosis of NSCLC cells. Luciferase assays and Western blotting were performed to validate potential targets of miR-654-3p. RESULTS: MiR-654-3p levels were significantly decreased in NSCLC patients and cell lines and were significantly correlated with the tumour size and tumour node metastasis stage of NSCLC patients. In A549 cells, miR-654-3p overexpression significantly increased apoptosis and inhibited growth both in vivo and in vitro, while downregulation of miR-654-3p had the opposite effects. In addition, polo-like kinase 4 (PLK4) was shown to be a target gene of miR-654-3p that is negatively regulated by miR-654-3p in A549 cells. Furthermore, PLK4 was observed to be highly expressed in NSCLC tissues and cells, and PLK4 overexpression abolished the inhibitory effects of miR-654-3p overexpression on NSCLC cell proliferation. Finally, the animal experiment results further demonstrated that miR-654-3p inhibits tumour growth and regulates PLK4 expression. CONCLUSION: Our results demonstrate that miR-654-3p functions as a growth-suppressing miRNA by targeting PLK4 in NSCLC.

Formation of highly ordered micro fillers in polymeric matrix by electro-field-assisted aligning.

Nanocomposites composed by polymeric matrix with micro/nano fillers have drawn lots of attention since their dramatic properties beyond pristine polymers. The spatial distribution of the micro/nano fillers in the polymeric matrix determines the final desired properties of the nanocomposites, thus deserves to investigate. Here, we proposed an effective method of assembling the micro/nano fillers to pre-designed patterns within the polymeric matrix by AC-electro-field-assisted aligning. By pre-designed AC electric fields which could be dynamically controllable, the distribution of microparticles (acting as fillers) in the matrix was tuned to various patterns related to the electric fields, such as linear alignment and circular alignment. The field-oriented particles chains could act as endoskeletal structures, showing unique properties (i.e., mechanical, optical, and anisotropic properties) beyond those of the conventional composites with randomly distributed particles.

Matrine induces apoptosis via targeting CCR7 and enhances the effect of anticancer drugs in non-small cell lung cancer in vitro.

This study mainly investigated the effects of matrine on cell apoptosis and the effects of anticancer drugs in non-small cell lung cancer (NSCLC) cell lines (A549 and LK2 cells). The results showed that matrine (≥10 µM) caused a significant inhibition on cell viability and 10 and 100 µM matrine induced cell apoptosis via influencing p53, bax, casp3, and bcl-2 expressions in A549 cells. In addition, matrine significantly down-regulated C-C chemokine receptor type 7 (CCR7) expression, and blocking the down-regulation of CCR7 by exogenous chemokine ligand 21 (CCL21) treatment alleviated matrine-caused effects of apoptosis genes in A549 cells. The results were further validated in LK2 cells that matrine regulated apoptosis gene expressions, which were reversed by CCL21 treatment. Furthermore, matrine enhances the effects of cisplatin, 5-fluorouracil, and paclitaxel in A549 cells, and the anticancer effects exhibit a dosage-dependent manner. In summary, matrine induced cell apoptosis and enhanced the effects of anticancer drugs in NSCLC cells; the mechanism might be associated with the CCR7 signal.