Metabolic and senescence characteristics associated with the immune microenvironment in non-small cell lung cancer: insights from single-cell RNA sequencing.

Non-small lung cancer (NSCLC) has been defined as a highly life-threatening heterogeneous disease, with high mortality and occurrence. Recent research has indicated that tumor-infiltrating lymphocytes play a key determinant role in cancer progression. Emerging single-cell RNA sequencing (also termed scRNA-seq) has been extensively applied to depict the baseline landscape of the cell composition and function phenotype in the tumor environment (TME). Herein, we dissected the cell types in NSCLC samples (including tissue and blood) and identified three types of cell marker genes including cancer cells, T cells, and macrophages by integrating two NSCLC-associated scRNA-seq datasets in GEO. Survival analysis indicated that 17 marker genes were related to tumor prognosis. Function annotation was used to scrutinize the molecular mechanism of these marker genes in different cells. Besides, we investigated the developmental trajectory and T cell receptor repertoire diversity of tumor-infiltrating T cells. Our analysis will help further understand the complexity of cell components and the heterogeneity of TME in NSCLC.

LncRNA ITGB2-AS1 promotes cisplatin resistance of non-small cell lung cancer by inhibiting ferroptosis via activating the FOSL2/NAMPT axis.

Cisplatin resistance is a major therapeutic challenge in non-small cell lung cancer (NSCLC). Herein, the regulatory role of long non-coding RNA (lncRNA) ITGB2-AS1 in regulating NSCLC cisplatin resistance was investigated. NSCLC cisplatin resistance cells were constructed using A549 and H1975 cells. Cell viability and proliferation were detected by MTT assay and colony formation assay, respectively. Cell apoptosis and cell cycle were examined by flow cytometry. GSH, MDA, ROS, and Fe2+ levels were measured by the corresponding kits. The expressions of ferroptosis-negative regulation genes (GPX4 and SLC7A11) were determined by qRT-PCR and western blot. Molecular interactions were analyzed by RNA pull-down, RIP, ChIP, and dual-luciferase reporter assays. The effects of ITGB2-AS1 silencing on NSCLC cisplatin resistance in vivo were elevated by the tumor xenograft experiment. ITGB2-AS1 expression was increased in NSCLC patients and cisplatin-resistant NSCLC cells, which was positively correlated with ferroptosis-negative regulation genes. ITGB2-AS1 knockdown suppressed resistant cell proliferation and promoted cell apoptosis and ferroptosis. ITGB2-AS1 increased NAMPT expression by binding to FOSL2, thereby repressing p53 expression. The ITGB2-AS1 knockdown also inhibited NSCLC cisplatin resistance in vivo. ITGB2-AS1 promoted NSCLC cisplatin resistance by inhibiting p53-mediated ferroptosis via activating the FOSL2/NAMPT axis.

Development of a 5-Gene Signature to Evaluate Lung Adenocarcinoma Prognosis Based on the Features of Cancer Stem Cells.

Cancer stem cells (CSCs) can induce recurrence and chemotherapy resistance of lung adenocarcinoma (LUAD). Reliable markers identified based on CSC characteristic of LUAD may improve patients' chemotherapy response and prognosis. OCLR was used to calculate mRNA expression-based stemness index (mRNAsi) of LUAD patients' data in TCGA. Association analysis of mRNAsi was performed with clinical features, somatic mutation, and tumor immunity. A prognostic prediction model was established with LASSO Cox regression. Kaplan-Meier Plotter (KM-plotter) and time-dependent ROC were applied to assess signature performance. For LUAD, univariate and multivariate Cox analysis was performed to identify independent prognostic factors. LUAD tissues showed a noticeably higher mRNAsi in than nontumor tissues, and it showed significant differences in T, N, M, AJCC stages, and smoking history. The most frequently mutated gene was TP53, with a higher mRNAsi relating to more frequent mutation of TP53. The mRNAsi was significantly negatively correlated with immune score, stromal score, and ESTIMATE score in LUAD. The blue module was associated with mRNAsi. The 5-gene signature was confirmed as an independent indicator of LUAD prognosis that could promote personalized treatment of LUAD and accurately predict overall survival (OS) of LUAD patients.

Mutational status of main driver genes influences the prognosis of stage I-III lung adenocarcinoma patients underwent radical surgery.

BACKGROUND: The therapeutic strategies and prognosis of local advanced and metastatic lung cancer have been extensively investigated. However, the prognosis of early-stage lung cancer patients undergoing radical surgery has not been fully studied due to the difficulties in follow-up and assessment. METHODS: We recruited 447 stage I-III lung adenocarcinoma (LUAD) patients who underwent radical surgery and investigated the influence of main driver gene mutations and clinicopathological factors on patient overall survival (OS). Cancer tissue samples were collected retrospectively and mutational status and tumor mutational burden (TMB) were determined by whole-exome sequencing (WES). RESULTS: Distinct stage-dependent mutational frequency was revealed in main driver genes including EGFR, TP53, KRAS, STK11, ATM and NF1. Patients with TP53 mutations exhibited a trend of better survival than those with wild type TP53 (P=0.066), and STK11 mutations exhibited worse survival in stage III patients (P=0.031). EGFR mutations eliminated the across-stage difference in survival, which was still present in other wild type and mutant driver genes. Furthermore, patients with wild type TP53 appeared to have significantly worse survival than patients with other wild type driver genes in stage I (P<0.001). TMB cannot stratify the survival of LUAD patients in stage I-III. Age, gender, smoking status, smoking years, prior cancer history and cancer location had no stratification effect on patient survival, while T grading (P<0.001) and N grading (P<0.001) had significant stratification on survival. CONCLUSIONS: TP53, EGFR and STK11 mutational status influenced the prognosis of stage I-III LUAD. T and N grading also stratified the patient survival. T grading was an independent risk factor.

miR­454­3p inhibits non­small cell lung cancer cell proliferation and metastasis by targeting TGFB2.

Accumulating studies have suggested that microRNAs (miRs) play a significant role in lung cancer development and progression, especially in non­small cell lung cancer (NSCLC). The present study aimed to investigate the associations between miR­454­3p and NSCLC progression. qPCR assay was applied to examine the expression of miR­454­3p and transforming growth factor­ß2 (TGFB2) in tissues and cell lines. CCK­8 and EdU assays were used to detect cell proliferation. Wound­healing and Transwell assays were conducted to assess cell migration and invasion. Western blotting assay was performed to explore the protein levels of epithelial­mesenchymal transition (EMT) markers. The interaction between miR­454­3p and TGFB2 was investigated with a luciferase reporter assay. miR­454­3p was downregulated in NSCLC tissues and NSCLC cell lines. miR­454­3p overexpression led to the suppression of proliferation, migration, and invasion in A549 and NCI­H1650 cells. In addition, the overexpression of miR­454­3p in A549 and NCI­H1650 cells significantly inhibited EMT. TGFB2 was revealed to be a direct target of miR­454­3p by using TargetScan database and luciferase reporter assay. TGFB2 was observed to be upregulated in NSCLC tissues and cell lines. Further mechanistic studies revealed that the inhibitory effects of miR­454­3p on NSCLC were reversed upon overexpression of TGFB2. These findings provided strong evidence that miR­454­3p suppressed NSCLC cell proliferation and metastasis by targeting TGFB2. The study suggests that targeting miR­454­3p could be a promising strategy for treating NSCLC.

[Retrospective study of clinicopathological characteristics in bronchopulmonary carcinoid].

BACKGROUND AND OBJECTIVE: Bronchopulmonary carcinoid (BPC) account for less than 2% of all primary lung malignant tumors, but few related studies were reported. The aim of this study is to analyze this rare disease's clinicopathological characteristics. METHODS: The clinical data of 28 patients with BPC in Cancer Center of Sun Yat-sen University, from January 1994 to June 2009, were enrolled into retrospective analysis. First, the corresponding paraffin blocks reexamined, slice up and stained, multiple pathologists re-consulted, and its subsets (typical carcinoid, TC; atypical carcinoid, AC) defined. Second, the clinical characteristics and immunohistochemical markers and its relationship with prognosis were analyzed. RESULTS: First, the 5-year survival for overall and TC, AC was 56% and 70%, 41% respectively in 28 cases. The markers CD99, Bcl-2 and Ki-67 expression correlated significantly with the BPC subsets (P = 0.017, P = 0.043, and P = 0.033 respectively). Further univariate analysis revealed that advanced TNM staging (P = 0.037), lymph node metastasis (P = 0.001) and Ki-67 nucleolus's positive expression (P = 0.009) are poor prognostic factors. Second, the overall, TC, AC 5-year survival rate was 73%, 83%, 57% respectively in 20 cases underwent the radically surgical resection. Further univariate analysis revealed that AC subset (P = 0.013), lymph node metastasis (P = 0.004) and Ki-67 nucleolus's positive expression (P = 0.006), advanced TNM staging (P = 0.047) are poor prognostic factors in this 20 cases. Third, as univariate analysis, local recurrence and metastasis (n = 4) correlate significantly with Ki-67 nucleolus's and Bcl-2 positive expression (P = 0.027, 0.045, respectively). CONCLUSION: The prognosis of BPC was better than other types of primary lung cancer. Ki-67, Bcl-2 high expression and advanced TNM staging are the poor recurrence and prognostic factors of BPC. The radical surgery remains the treatment of choice for resectable candidates in BPC as NSCLC.