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.

Superior improvement on soil quality by Pennisetum sinese vegetation restoration in the dry-hot valley region, SW China.

Vegetation restoration is a good way to improve soil quality and reduce erosion. However, the impact of vegetation restoration on soil quality in the dry-hot valley region has been overlooked for many years. This study aimed to reveal the effects of Pennisetum sinese (PS) and natural vegetation (NV) on soil quality and then to explore the feasibility of introducing PS for the vegetation restoration of the dry-hot valley region. The PS and NV restoration areas deserted land evolving from cultivated land (CL) have been established since 2011. The results showed that the soil properties were obviously improved by PS from the dry to wet seasons, except for the soil available phosphorous. The comprehensive soil quality indexes of the three typical seasons (dry, dry-wet, and wet) were determined by using nonlinear weighted additive (NLWA) based on the total dataset, significant dataset and minimum dataset (MDS). The results indicated that the comprehensive minimum dataset soil quality index (MDS-SQI) of the three typical seasons evaluate soil quality well. The soil quality of PS was significantly greater than that of CL and NV (P < 0.05), as shown by the MDS-SQI. Additionally, PS could maintain a stable soil quality in the three typical seasons, while both CL and NV had obvious fluctuations. In addition, the result of the generalized linear mode suggested that the vegetation type had the greatest impact on the soil quality (44.51 %). Comprehensively, vegetation restoration in the dry-hot valley region has a positive impact on the soil properties and quality. PS is a great candidate species for the early vegetation restoration in the dry-hot valley region. This work provides a reference for vegetation restoration and rational utilization of soil resources in degraded ecosystems in dry-hot valleys and other soil erosion areas.

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.

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.

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.