Non-small Cell Lung Cancer Lung Research Articles

PLA2G10 incorporated in exosomes could be diagnostic and prognostic biomarker for non-small cell lung cancer

BackgroundExosomal cargos such as nucleic acids and proteins have been attracting major interest as promising diagnostic biomarkers of cancers. The aim of this study was to characterize the mRNA profiles of serum exosomes and to identify non-small cell lung cancer (NSCLC) related mRNAs with higher sensitivity and specificity to diagnose and predict prognosis of NSCLC. MethodsmRNA microarray analysis was conducted to screen differentially expressed mRNAs in the serum exosomes of NSCLC patients. Selected exosomal mRNA candidate PLA2G10 and PLA2G10 protein were quantified by RT-qPCR and ELISA assay, respectively, in the sample cohorts of healthy, benign lung tumor and NSCLC. Receiver operating characteristic (ROC) analyses were performed to evaluate the diagnostic power of exosomal PLA2G10 mRNA and protein. Kaplan-Meier plots were used to estimate patients’ overall and disease-free survival. ResultsSerum exosomal PLA2G10 mRNA levels were elevated in NSCLC patients, and were closely related to more aggressive characteristics (higher stages, lymphatic node metastasis and distant metastasis) and poor overall and disease-free survival of NSCLC patients. Intriguingly, PLA2G10 protein was proved to be incorporated in exosomes, and its expression patterns and relationship with clinical pathological factors were similar to exosomal PLA2G10 mRNA. Additionally, the levels of exosomal PLA2G10 mRNA and protein were positively correlated and their combination could improve the diagnostic power to discriminate less and more malignance of NSCLC. ConclusionsIncreased levels of serum exosomal PLA2G10 mRNA and protein were associated with more aggressive features of NSCLC, suggesting their potential as diagnostic and prognostic biomarkers of NSCLC.

IDO1 can impair NK cells function against non-small cell lung cancer by downregulation of NKG2D Ligand via ADAM10

Indoleamine 2,3-dioxygenase 1 (IDO1) catalyzes the rate-limiting step in tryptophan catabolism along the kynurenine (Kyn) pathway and exerts immunosuppressive properties mainly via activation of transcription factor aryl hydrocarbon receptor (AhR) pathway. IDO1 induces NK cells dysfunction via downregulation of the activating receptor NKG2D on NK cells, but whether and how it affects the expression of NKG2D Ligand (NKG2DL) on tumor cells remains unclear. Since a disintegrin and metalloprotease 10 (ADAM10) plays a potential role in the shedding of NKG2DL and the releasing of soluble NKG2DL (sNKG2DL), we investigated how IDO1 modulates the expression of NKG2DL via ADAM10 in non-small cell lung cancer (NSCLC). We found that IDO1 expression was negatively correlated with NKG2DL expression while positively correlated with ADAM10 expression with human lung cancer brain metastasis tissue, NSCLC cells and LLC tumor-bearing mice. IDO1 could regulate ADAM10 expression via IDO1-Kyn-AhR signaling pathway and subsequently regulate NKG2DL expression. IDO1 deficiency led to retarded tumor growth and improved NK cells function in NSCLC mice. IDO1 inhibitors improved NK cells function in vitro and in vivo. The combo of IDO1 inhibitor and NK cells exhibited more therapeutic efficacy than either of the single IDO1 inhibitor or NK cells treatment.

CircCAMSAP1 promotes non-small cell lung cancer proliferation and inhibits cell apoptosis by sponging miR-1182 and regulating BIRC5

ABSTRACT Recently, various studies have suggested that circular RNAs (circRNAs) are ubiquitous in various malignant events, including non-small cell lung cancer (NSCLC) and are closely related to cell proliferation and apoptosis. Unfortunately, the molecular functions involved in this action still have little overlap. Therefore, this study aimed to identify a novel circCAMSAP1 role in NSCLC. Overexpression of circCAMSAP1 has been demonstrated in NSCLC lung tissues and cell lines. Sequencing and RNase R experiments were planned to determine whether circCAMSAP1 is looped and exists in NSCLC. We also found that downregulated circCAMSAP1 repressed cell proliferation and increased apoptosis of NSCLC cells in vitro and suppressed xenograft tumor growth in vivo. Furthermore, a luciferase assay revealed that circCAMSAP1 could regulate baculoviral inhibitor of apoptosis protein (IAP) repeat containing 5 (BIRC5, also known as survivin) expression by directly binding to miR-1182. However, BIRC5 without 3ʹ untranslated regions (3ʹUTR) could reverse the influence of downregulated circCAMSAP1 on proliferation and apoptosis in NSCLC. Together, our findings reveal a novel mechanism by which the circCAMSAP1/miR-1182/BIRC5 axis promotes NSCLC progression.

Multimodal analysis suggests differential immuno-metabolic crosstalk in lung squamous cell carcinoma and adenocarcinoma

Immunometabolism within the tumor microenvironment is an appealing target for precision therapy approaches in lung cancer. Interestingly, obesity confers an improved response to immune checkpoint inhibition in non-small cell lung cancer (NSCLC), suggesting intriguing relationships between systemic metabolism and the immunometabolic environment in lung tumors. We hypothesized that visceral fat and 18F-Fluorodeoxyglucose uptake influenced the tumor immunometabolic environment and that these bidirectional relationships differ in NSCLC subtypes, lung adenocarcinoma (LUAD) and lung squamous cell carcinoma (LUSC). By integrating 18F-FDG PET/CT imaging, bulk and single-cell RNA-sequencing, and histology, we observed that LUSC had a greater dependence on glucose than LUAD. In LUAD tumors with high glucose uptake, glutaminase was downregulated, suggesting a tradeoff between glucose and glutamine metabolism, while in LUSC tumors with high glucose uptake, genes related to fatty acid and amino acid metabolism were also increased. We found that tumor-infiltrating T cells had the highest expression of glutaminase, ribosomal protein 37, and cystathionine gamma-lyase in NSCLC, highlighting the metabolic flexibility of this cell type. Further, we demonstrate that visceral adiposity, but not body mass index (BMI), was positively associated with tumor glucose uptake in LUAD and that patients with high BMI had favorable prognostic transcriptional profiles, while tumors of patients with high visceral fat had poor prognostic gene expression. We posit that metabolic adjunct therapy may be more successful in LUSC rather than LUAD due to LUAD’s metabolic flexibility and that visceral adiposity, not BMI alone, should be considered when developing precision medicine approaches for the treatment of NSCLC.

Bone metastases tend to increase in non-small cell lung cancer with epidermal growth factor receptor mutation

BackgroundIncreased understanding in molecular pathology of advanced non-small cell lung cancer (NSCLC) over the past decades has led to personalized treatment approaches being advocated. Epidermal growth factor receptor (EGFR) mutation that often occurs in NSCLC can be identified using immunohistochemical examinations. Moreover, clarifying the relationship between computed tomography (CT) and EGFR mutation of NSCLC might inform therapeutic decision-making. The purpose of this study was to determine the relationship between metastatic sites on primary chest CT-scan and EGFR mutation in NSCLC lung cancer patients. MethodsAn cross-sectional design using secondary data was conducted, involving 76 NSCLC patients. EGFR mutations were determined by immunohistochemical examination and metastatic sites by chest CT-scan with contrast. The collected metastatic sites comprised hilar and mediastinal lymphadenopathy, pulmonary nodules, and bone, liver, spleen and suprarenal metastases. A Chi square test was used to analyze the data. ResultsThis study revealed that the highest NSCLC stage was IVb, found in 39 samples (51.3%), while 34 (44.7%) subjects had EGFR mutation. There was no statistically significant difference between metastatic site and positive EGFR mutation, although positive bone metastases (54.8%) tend to have more numerous positive EGFR mutations compared to negative bone metastases (37.7%) (p=0.142). ConclusionsPatients with positive bone metastases tend to have higher positive EGFR mutation compared to negative bone metastases in NSCLC lung cancer patients. Prospective studies evaluating patients with EGFR mutation for bone metastases should be considered. This can provide information on therapeutic decision-making to obtain good clinical outcomes.

Serum exosomal hsa_circ_0069313 has a potential to diagnose more aggressive non-small cell lung cancer

BackgroundCircular RNAs (circRNAs) derived from exosomes are involved in the carcinogenesis and development of non-small cell lung cancer (NSCLC), showing great potential to be diagnostic biomarkers for NSCLC. MethodsSerum exosomes were isolated with an exosome isolation kit, and verified by western blot, transmission electron microscopy and a potentiometric analyzer. Five differentially expressed exosomal circRNAs, including hsa_circ_0069313, hsa_circ_0063526, hsa_circ_0010522, hsa_circ_0048677 and hsa_circ_0001946, were selected based on the circRNA array analyses and the published documents in Pubmed. The serum and serum exosomal levels of the above five circRNAs were quantified by qRT-PCR. The diagnostic power of serum and serum exosomal hsa_circ_0069313 was evaluated by receiver operating characteristic (ROC) test. ResultsThe levels of hsa_circ_0069313 in serum exosomes were statistically lower than those in the matched serum samples. In contrast, the levels of hsa_circ_0063526, hsa_circ_0010522, hsa_circ_0048677 and hsa_circ_0001946 showed no statistical difference in the sera and serum exosomes of healthy donors. The levels of serum and serum exosomal hsa_circ_0069313 were notably elevated in the NSCLC group compared to the healthy, pneumonia and benign lung tumor groups. Furthermore, serum and serum exosomal hsa_circ_0069313 could differ benign lung tumor and NSCLC with AUC values of 0.803 and 0.749, respectively. Intriguingly, the higher levels of serum exosomal hsa_circ_0069313 were associated with stage III-IV, lymph node metastasis and distant metastasis of NSCLC. ConclusionsSerum and serum exosomal hsa_circ_0069313 have the potential to discriminate NSCLC and benign lung tumor. The higher levels of serum exosomal hsa_circ_0069313 are linked to more aggressive pathological features of NSCLC.

MiR-421 promotes lipid metabolism by targeting PTEN via activating PI3K/AKT/mTOR pathway in non-small cell lung cancer.

Aims: We aim to investigate the effects of miR-421 on lipid metabolism in non-small cell lung cancer (NSCLC). Methods: The miR-421 expression and PTEN mRNA level in tumor tissues, adjacent normal tissues, human lung epithelial cells and NSCLC cell lines were detected with reverse transcription quantitative real-time PCR. Results: MiR-421 was increased, and PTEN was reduced remarkably in tumor tissues and NSCLC cell lines. Down-regulated miR-421 suppressed lipid accumulation, cell proliferation, migration and invasion, whereas overexpression of miR-421 had the opposite effects. MiR-421 directly targeted PTEN and negatively regulated PTEN expression. MiR-421 activated PI3K/AKT/mTOR pathway through regulating PTEN. Conclusion: MiR-421 promotes lipid metabolism through targeting PTEN via PI3K/AKT/mTOR pathway activation in NSCLC, indicating that miR-421 can be a latent therapeutic target for NSCLC.

MicroRNA-1321 and microRNA-7515 contribute to the progression of non-small cell lung cancer by targeting CDC20.

Cell division cycle 20 (CDC20) and microRNAs (miRNAs) are differentially expressed in non-small cell lung cancer (NSCLC). The current study aimed to investigate the role of miR-1321 and miR-7515 regulation in CDC20 during NSCLC development. CDC20 expression in paracancerous and tumor tissues was assessed using quantitative reverse transcription-polymerase chain reaction (qRT-PCR). The relationship between CDC20 expression and prognosis of patients was analyzed using the TCGA database. The expression profile of CDC20 in healthy lung cells and NSCLC cells was detected using qRT-PCR and western blotting. After the knockdown of CDC20 in NSCLC cells, the cell proliferation, apoptosis, migration, invasion, and cell cycle changes were investigated by CCK8, EdU, flow cytometry, wound healing, and Transwell assays. The miRNAs targeting CDC20 were predicted using two bioinformatics websites and validated using dual-luciferase assays. CDC20 was enhanced in NSCLC tissues and cells, thus predicting the poor prognosis in NSCLC patients. After CDC20 inhibition, the malignant phenotype of NSCLC cells was reverted. miR-1321 and miR-7515 targeted CDC20 and exhibited the same anti-tumor effects as CDC20 silencing. Functional rescue experiments showed that CDC20 overexpression averted the anti-tumor effects of miR-1321 and miR-7515 on NSCLC cells. miR-1321 and miR-7515 inhibited NSCLC development by targeting CDC20. Thus, the current study has implications in NSCLC treatment and provides novel insights into NSCLC management.

A Signature of 14 Long Non-Coding RNAs (lncRNAs) as a Step towards Precision Diagnosis for NSCLC.

Simple SummaryAlthough the biological function of lncRNAs has not been fully elucidated, we know that the aberrant expression of lncRNAs can drive the cancer phenotype. Therefore, a growing area of research is focusing on lncRNAs as putative diagnostic biomarkers and therapeutic targets. The aim of the study was the appraisal of the diagnostic value of 14 differentially expressed lncRNA in the early stages of NSCLC. We established two classifiers. The first recognized cancerous from noncancerous tissues, the second successfully discriminated NSCLC subtypes (LUAD vs. LUSC). Our results indicate that the panel of 14 lncRNAs can be a promising tool to support a routine histopathological diagnosis of NSCLC.LncRNAs have arisen as new players in the world of non-coding RNA. Disrupted expression of these molecules can be tightly linked to the onset, promotion and progression of cancer. The present study estimated the usefulness of 14 lncRNAs (HAGLR, ADAMTS9-AS2, LINC00261, MCM3AP-AS1, TP53TG1, C14orf132, LINC00968, LINC00312, TP73-AS1, LOC344887, LINC00673, SOX2-OT, AFAP1-AS1, LOC730101) for early detection of non-small-cell lung cancer (NSCLC). The total RNA was isolated from paired fresh-frozen cancerous and noncancerous lung tissue from 92 NSCLC patients diagnosed with either adenocarcinoma (LUAD) or lung squamous cell carcinoma (LUSC). The expression level of lncRNAs was evaluated by a quantitative real-time PCR (qPCR). Based on Ct and delta Ct values, logistic regression and gradient boosting decision tree classifiers were built. The latter is a novel, advanced machine learning algorithm with great potential in medical science. The established predictive models showed that a set of 14 lncRNAs accurately discriminates cancerous from noncancerous lung tissues (AUC value of 0.98 ± 0.01) and NSCLC subtypes (AUC value of 0.84 ± 0.09), although the expression of a few molecules was statistically insignificant (SOX2-OT, AFAP1-AS1 and LOC730101 for tumor vs. normal tissue; and TP53TG1, C14orf132, LINC00968 and LOC730101 for LUAD vs. LUSC). However for subtypes discrimination, the simplified logistic regression model based on the four variables (delta Ct AFAP1-AS1, Ct SOX2-OT, Ct LINC00261, and delta Ct LINC00673) had even stronger diagnostic potential than the original one (AUC value of 0.88 ± 0.07). Our results demonstrate that the 14 lncRNA signature can be an auxiliary tool to endorse and complement the histological diagnosis of non-small-cell lung cancer.

Identification of SRXN1 and KRT6A as Key Genes in Smoking-Related Non-Small-Cell Lung Cancer Through Bioinformatics and Functional Analyses.

BackgroundLung cancer is the leading cause of cancer-related mortality worldwide. Although cigarette smoking is an established risk factor for lung cancer, few reliable smoking-related biomarkers for non-small-cell lung cancer (NSCLC) are available. An improved understanding of these biomarkers would further the development of new biomarker-targeted therapies and lead to improvements in overall patient survival.MethodsWe performed bioinformatic analysis to screened potential target genes, then quantitative PCR, western, siRNA, CCK-8, flow cytometry, tumorigenicity assays in nude mice were performed to validated the function.ResultsIn this study, we identified 83 smoking-related genes (SRGs) based on an integration analysis of two Gene Expression Omnibus (GEO) datasets, and 27 hub SRGs with potential carcinogenic effects by analyzing a dataset of smokers with NSCLC in The Cancer Genome Atlas (TCGA) database. A survival analysis revealed three genes with potential prognostic value, namely SRXN1, KRT6A and JAKMIP3. A univariate Cox analysis revealed significant associations of elevated SRXN1 and KRT6A expression with prognosis. A receiver operating characteristic (ROC) curve analysis indicated the high diagnostic value of SRXN1 and KRT6A for smoking and cancer. Quantitative PCR and western blotting validated the increased expression of SRXN1 and KRT6A mRNA and protein, respectively, in lung cancer cell lines and NSCLC tissues. In patients with NSCLC, SRXN1 and KRT6A expression was associated with the tumor–node–metastasis (TNM) stage, presence of metastasis, history of smoking and daily smoking consumption. Furthermore, inhibition of SRXN1 or KRT6A suppressed viability and enhanced apoptosis in the A549 human lung carcinoma cell line. Tumorigenicity assays in nude mice confirmed that the siRNA-mediated downregulation of SRXN1 and KRT6A expression inhibited tumor growth in vivo.ConclusionsIn summary, SRXN1 and KRT6A act as oncogenes in NSCLC and might be potential biomarkers of smoking exposure and the early diagnosis and prognosis of NSCLC in smokers, which is vital for lung cancer therapy.

Лечение немелкоклеточного рака легкого с использованием лучевой терапии

The article is devoted to the use of a combination of systemic and radiation therapy in patients with locally advanced non-small cell lung cancer (NSCLC). The high risk of latent metastasis in this category of patients sets the prerequisites for active research of the combination of radiation therapy and systemic treatment worldwide. The article presents the results of clinical studies aimed to find the most optimal ratio of efficacy and safety of the chemotherapeutic component of chemoradiotherapy (CRT). According to the results of the research, the authors prefer the use of schemes based on platinum doublets. The article discusses the evaluation of the effectiveness of chemoradiotherapy with escalation of the radiation dose to the tumor within the framework of the concept of a personalized approach to therapy of patients with NSCLC. The article also pays attention to the description of new possibilities for RT intensification and the introduction of new RT methods, the use of which became possible with the introduction of innovative linear accelerators. The results of comparing the effectiveness of combined chemoradiotherapy with immunotherapy and chemo radiotherapy alone in patients with lung NSCLC are presented.

A MOLECULAR DOCKING STUDY OF EPOXYXANTHONES TOWARD NON-SMALL AND SMALL CELL LUNG CANCER TARGET PROTEINS

Molecular docking studies have been used to evaluate the anticancer activity of the epoxyxanthones against non-small cell lung cancer (NSCLC) and small cell lung cancer (SCLC). A series of twenty-four xanthone compounds possessing epoxy and hydroxy substituents have been designed and investigated. The epoxyxanthone derivatives were docked in the active site of EGFR and KIT tyrosine kinase proteins to examine their inhibition activity toward two types of lung cancer, i.e., NSCLC and SCLC, respectively. The xanthone derivatives were successfully docked in a similar binding mode to native ligands on EGFR and KIT proteins with free binding energies ranging from -7.51 to -6.35 and -9.58 to -7.06 kcal/mol, respectively. It was found that compound XE9 exhibited the most promising activity with a free binding energy value of -9.58 and -7.51 kcal/mol against KIT and EGFR proteins, respectively. This study discovered that XE9 has potency as a therapeutic agent used as the SCLC and NSCLC inhibitors or used in the combination of the cells found in lung cancer.

SchizandrinA can inhibit non‑small cell lung cancer cell proliferation by inducing cell cycle arrest, apoptosis and autophagy.

Schizandrin A (SchA) can be extracted from the vine plant Schisandra chinensis and has been reported to confer various biologically active properties. However, its potential biological effects on non-small cell lung cancer (NSCLC) remain unknown. Therefore, the present study aims to address this issue. NSCLC and normal lung epithelial cell lines were first treated with SchA. Cell viability and proliferation were measured using CellTiter-Glo Assay and colony formation assays, respectively. PI staining was used to measure cell cycle distribution. Cell cycle-related proteins p53, p21, cyclin D1, CDK4, CDK6, cyclin E1, cyclin E2, CDK2 and DNA damage-related protein SOX4 were detected by western blot analysis. Annexin V-FITC/PI staining, DNA electrophoresis and Hoechst 33342/PI dual staining were used to detect apoptosis. JC-1 and DCFH-DA fluorescent dyes were used to measure the mitochondrial membrane potential and reactive oxygen species concentrations, respectively. Apoptosis-related proteins caspase-3, cleaved caspase-3, poly(ADP-ribose) polymerase (PARP), cleaved PARP, BimEL, BimL, BimS, Bcl2, Bax, caspase-9 and cleaved caspas-9 were measured by western blot analysis. Dansylcadaverine was used to detect the presence of the acidic lysosomal vesicles. The expression levels of the autophagy-related proteins LC3-I/II, p62/SQSTM and AMPKα activation were measured using western blot analysis. In addition, the autophagy inhibitor 3-methyladenine was used to inhibit autophagy. SchA treatment was found to reduce NSCLC cell viability whilst inhibiting cell proliferation. Low concentrations of SchA (10-20 µM) mainly induced G1/S-phase cell cycle arrest. By contrast, as the concentration of SchA used increases (20-50 µM), cells underwent apoptosis and G2/M-phase cell cycle arrest. As the treatment concentration of SchA increased from 0 to 50 µM, the expression of p53 and SOX4 protein also concomitantly increased, but the expression of p21 protein was increased by 10 µM SchA and decreased by higher concentrations (20-50 µM). In addition, the mRNA and protein expression levels of Bcl-like 11 (Bim) EL, BimL and BimS increased following SchA application. SchA induced the accumulation of acidic vesicles and induced a marked increase in the expression of LC3-II protein, suggsting that SchA activated the autophagy pathway. However, the expression of the p62 protein was found to be increased by SchA, suggesting that p62 was not degraded during the autophagic flux. The 3-methyladenine exerted no notable effects on SchA-induced apoptosis. Taken together, results from the present study suggest that SchA exerted inhibitory effects on NSCLC physiology by inducing cell cycle arrest and apoptosis. In addition, SchA partially induced autophagy, which did not result in any cytoprotective effects.

Survival rates of patients with tumors originating in different segments of the left upper lung in stage I to III non-small cell lung cancer.

BackgroundThe aim of this research was to evaluate the effect of spatial location of tumors on the prognosis of patients with left upper lung non-small cell lung cancer (NSCLC), with a focus on the S1+2+3 and lingual segment.MethodsA total of 486 patients who underwent lobectomy and systematic lymph node dissection were collected retrospectively in this study (354 S1+2+3 and 132 lingual segment patients). Factors impacting survival were assessed via univariate analyses, multivariate analyses, and log-rank tests.ResultsCompared with tumor location in S1+2+3, lingual segment tumor location of stage II to III left upper lung NSCLC patients was significantly associated with a better 5-year disease-free survival (DFS) (P=0.041). Multivariate analysis results showed that tumor location in the lingual segment was a good independent prognostic factor of stage II to III left upper lung NSCLC patients [hazard ratio (HR) =0.602, 95% confidence interval (CI): 0.149–0.865, P=0.006). However, in stage I left upper lung NSCLC, tumor location (HR =1.069, 95% CI: 0.571–2.000, P=0.835) was not an independent prognostic factor, and only T2 (HR =2.422, 95% CI: 1.271–4.620, P=0.007) was an independent worse prognosis factor.ConclusionsTumor location in the lingual segment of left upper lung stage II to stage III NSCLC is a good independent prognostic factor compared with S1+2+3. Nevertheless, tumor location does not impact the prognosis of patients with stage I NSCLC in the left upper lung.

LncRNA cytoskeleton regulator reduces non‑small cell lung cancer radiosensitivity by downregulating miRNA‑206 and activating prothymosin α.

The present study aimed to explore the role of the long noncoding RNA cytoskeleton regulator (CYTOR) in non-small cell lung cancer (NSCLC) radiosensitivity by manipulating the microRNA (miR)-206/prothymosin α (PTMA) axis. First, 58 pairs of NSCLC and paracancerous tissues, normal human lung epithelial cells and NSCLC cells were collected to analyze CYTOR expression and the relation- ship between CYTOR and NSCLC prognosis. Subsequently, CYTOR expression in radioresistant cells was assessed. Radioresistant cells with low CYTOR expression and parental cells with high CYTOR expression were established. Functional assays were then performed to assess changes in cell radiosensitivity after irradiation treatment. Subsequently, the downstream mechanism of CYTOR was explored. The binding interactions between CYTOR and miR-206 and between miR-206 and PTMA were predicted and certified. Xenograft transplantation was applied to confirm the role of CYTOR in the radiosensitivity of NSCLC. CYTOR was overexpressed in NSCLC and was associated with poor prognosis. CYTOR was further upregulated in NSCLC cells with radioresistance. CYTOR knockdown enhanced the radiosensitivity of NSCLC cells, while overexpression of CYTOR led to the opposite result. Mechanistically, CYTOR specifically bound to miR-206 and silencing CYTOR promoted miR-206 to enhance the radiosensitivity of NSCLC cells. PTMA is a target of miR-206 and silencing CYTOR inhibited PTMA expression via miR-206, thus promoting radiosensitivity of NSCLC cells. CYTOR knockdown also enhanced NSCLC cell radiosensitivity in vivo. CYTOR was highly expressed in NSCLC, while silencing CYTOR potentiated NSCLC cell radiosensitivity by upregulating miR-206 and suppressing PTMA. The present study preliminarily revealed the role of CYTOR in radiotherapy sensitivity of NSCLC and provided a novel potential target for the clinical treatment of NSCLC.

FAIM2 Promotes Non-Small Cell Lung Cancer Cell Growth and Bone Metastasis by Activating the Wnt/β-Catenin Pathway.

AimBone metastasis is the major reason for the poor prognosis and high mortality rate of non-small cell lung cancer (NSCLC) patients. This study explored the function and underlying mechanism of Fas apoptotic inhibitory molecule 2 (FAIM2) in the bone metastasis of NSCLC.MethodsSamples of normal lung tissue and NSCLC tissue (with or without bone metastasis) were collected and analyzed for FAIM2 expression. HARA cells with FAIM2 overexpression and HARA-B4 cells with FAIM2 knockdown were tested for proliferation, migration, invasion, anoikis, and their ability to adhere to osteoblasts. Next, whether FAIM2 facilitates bone metastasis by regulating the epithelial mesenchymal transformation (EMT) process and Wnt/β-catenin signaling pathway were investigated. Finally, an in vivo model of NSCLC bone metastasis was established and used to further examine the influence of FAIM2 on bone metastasis.ResultsFAIM2 was highly expressed in NSCLC tissues and NSCLC tissues with bone metastasis. FAIM2 expression was positively associated with the tumor stage, lymph node metastasis, bone metastasis, and poor prognosis of NSCLC. FAIM2 upregulation promoted HARA cell proliferation, migration, and invasion, but inhibited cell apoptosis. FAIM2 knockdown in HARA-B4 cells produced the opposite effects. HARA-B4 cells showed a stronger adhesive ability to osteocytes than did HARA cells. FAIM2 was found to be related to the adhesive ability of HARA and HARA-B4 cells to osteocytes. FAIM2 facilitated bone metastasis by regulating the EMT process and Wnt/β-catenin signaling pathway. Finally, FAIM2 was found to participate in regulating NSCLC bone metastasis in vivo.ConclusionsFAIM2 promoted NSCLC cell growth and bone metastasis by regulating the EMT process and Wnt/β-catenin signaling pathway. FAIM2 might be useful for diagnosing and treating NSCLC bone metastases.

Traitements par radiothérapie stéréotaxique : indications et situations cliniques particulières: Stereotactic radiotherapy: Indications and specific clinical settings

Lobectomy with radical lymph node dissection remains the gold standard treatment for stage I primary non-small cell lung cancer (NSCLC) but in recent years, stereotaxic radiotherapy (SBRT) have gradually taken their place in this indication. The choice of doses and fractionation and the stereotactic radiotherapy technique should be considered taking into account the patient’s clinical situation and the tumour to be treated (size and topography.)Some specials clinical setting as the treatment of ground glass opacity remain questionable without a lot of evidence in scientifics publications but offer a real alternative of treatment for inoperables patients. In regards to catch-up treatments, reirradiations offer new perspectives in case of local failure after stereotactic radiotherapy with promising results. But the majority of relapses are regional or metastatic and systemic treatment especially adjuvant immunotherapy is a very interesting treatment option for this frail patients, not enough fit to receive chemotherapy.The spontaneous evolution of NSCLC lung tumors without treatment remain very negative; The use of local non-surgical treatments as SBRT is an major option for the care of elderly patients who are not eligible for surgery. Despite their comorbidities, frailty and their poor remaining survival the majority of this patients will die of their lung cancer.© 2021 SPLF. Published by Elsevier Masson SAS. All rights reserved.

LncRNA OGFRP1 acts as an oncogene in NSCLC via miR-4640-5p/eIF5A axis

BackgroundLong noncoding RNAs (lncRNAs) OGFRP1 is up-regulated in endometrial cancer and cervical carcinoma, and OGFRP1 suppression inhibits the malignant behavior of cancer cells. Here, we evaluated the expression pattern, biological function and potential mechanism of OGFRP1 in non-small cell lung cancer (NSCLC).MethodsThe expression of target genes in 25 pairs of clinically collected NSCLC and normal lung tissue samples was detected by qRT-PCR or western blot. We screened the siRNA (siOGFRP1) to down-regulate the expression of OGFRP1 in A549 and H1299 cells. The biological function of A549 and H1299 cells were examined by CCK8, wound healing and transwell assays. The molecular mechanism of OGFRP1 was further explored.ResultsThe expression of OGFRP1 in NSCLC tissues were higher than that in normal lung tissue. siOGFRP1 inhibited the proliferation, migration and invasion of A549 and H1299 cells. In addition, the expression of EMT-related and apoptosis-related proteins was changed by siOGFRP1 transfection. OGFRP1 can directly interact with miR-4640-5p, and siOGFRP1 increased the level of miR-4640-5p. Moreover, miR-4640-5p could directly bind to the 3’ UTR region of eIF5A mRNA. eIF5A was highly expressed in NSCLC tissues, and predicted a poor prognosis. In addition, the expression of miR-4640-5p and eIF5A in NSCLC tissues were negatively correlated, while the expression of OGFRP1 and eIF5A were positively correlated. Knockdown of OGFRP1 inhibited the expression of eIF5A, while transfection of miR-4640-5p inhibitor up-regulated the expression of eIF5A.ConclusionsTaken together, we demonstrated that down-regulation of OGFRP1 inhibited the progression of NSCLC through miR-4640-5p/eIF5A axis.

Targeting DNAJC19 overcomes tumor growth and lung metastasis in NSCLC by regulating PI3K/AKT signaling

BackgroundSome driver oncogenes are still unknown in non-small-cell lung cancer (NSCLC). DNAJC19, a major component of the translocation machinery of mitochondrial membranes, is a disease-associated protein. Herein, we report the role of DNAJC19 in NSCLC cell growth and metastasis.MethodsImmunohistochemistry (IHC) was performed to investigate DNAJC19 expression in NSCLC clinical samples. For knockdown or overexpression assays in A549 or NCI-H1299 lung cancer cells, lentiviral vectors were used. After assessment of cell functions, DNAJC19-knockdown A549 cells were further applied to establish mouse xenograft and metastasis tumor models. Assessments based on the RNA-seq data, western blotting, PCR and IHC were performed for the mechanistic study.ResultsExpression of DNAJC19 was higher in tumors than in noncancerous adjacent tissues. Survival analysis indicated that low DNAJC19 levels were correlated with an increased progression-free survival rate. ShRNA-mediated knockdown of DNAJC19 markedly inhibited cell growth, viability, migration and invasion. Moreover, RNA-seq analysis revealed that the PI3K/AKT signaling pathway was involved in molecular events when A549 cells were treated with shDNAJC19. In addition, DNAJC19 knockdown decreased PI3Kp85a, AKT and p-AKT expression in A549 cells, and cellular functions were greatly rescued in DNAJC19-knockdown A549 cells by ectopic overexpression of AKT. Furthermore, tumor xenograft growth and lung metastasis were markedly repressed in the shDNAJC19 group compared to the control group. As expected, the expression levels of DNAJC19, PI3K and AKT in xenograft mouse samples were also lower in the shDNAJC19 group than in the shCtrl group.ConclusionsDNAJC19 greatly promotes NSCLC cell growth and lung metastasis by regulating PI3K/AKT signaling, providing a novel therapeutic target for treating NSCLC patients.

Abstract 618: Spatio-temporal multiomics analysis reveals distinct molecular features in recurrent stage I non-small cell lung cancers after R0 tumor resection

Abstract A portion of stage I non-small cell lung cancer (NSCLC) patients who have received R0 resection experience unexpected tumor recurrence during follow up. Some of them even recurred within 12 months after surgery, which is unable to predict by any clinical characteristics. Few study investigates the molecular features of such short-term recurrent tumors. Multiomics analysis were performed on 81 stage I NSCLC patients who had received R0 resection, based on deep whole-exome sequencing (average depth >500x) on multi-region 239 tumor tissue samples and 81 matched-adjacent normal specimens, as well as RNA sequencing and plasma targeted circulating tumor DNA detection. Dynamic dimension comparison of genomic alteration features between tumor recurrence and non-recurrence patients was made according to 60-months median time of follow up. Published WES datasets of Stage I NSCLCs of 131 East Asian (EAS LUADs), 62 European (TRACERx) and 277 American patients (TCGA) were extracted as validation. 60.5% patients were non-smokers. Lower chromosomal instability was found in our cohort characterized by less trunk CNVs and lower CNV coverage on genome as compared to the European cohort. Age-related signature mutations significantly accumulated in recurrence-free tumors. Somatic mutations occurred in DNA damage repair pathways were more likely to cause tumor recurrence (p=0.04), particularly in homologous recombination (HRR) (p=0.017) and translesion synthesis (TLS) (p=0.009) pathways. Dynamic analysis discovered the essential genes in Fanconi anemia (FA), TLS and HRR pathways that coordinate together for DNA interstrand crosslink (ICL) recognition and faithful repair of DNA double strand breaks (DSBs) during DNA replication stress frequently mutated in early tumor recurrence patients (n=7/11, 63%). Twenty-two focal CNV-driver genes, including cell proliferation-related RECQL4 (8q24.3), PIK3CA (3q26.32), BCL9 (1q21.2), TBL1XR1 (3q26.32) frequently amplified in recurrent tumors, as well as BCL3 and CBLC (19q13.31-32) in the early recurrent tumors. RNA-seq data demonstrated cell cycle pathway significantly up-regulated in recurrent tumors. More TMB, CNV fraction and genome doubling events in FA-TLS-HRR alteration patients verified genomic instability caused by dysfunction of cell cycle control and ICL-DSB repair contributing to early tumor recurrence. Integrative model cross clinical and molecular features stratified stage I patient prognosis in both our cohort and European cohort, in which and high risk patients were more detected by circulating tumor DNA. This is the first study to dynamically investigate the genomic features specifically in stage I lung NSCLC according to the precise tumor recurrence time, which provides important clues to postoperative treatment strategy and drug target research. Citation Format: Kezhong Chen, Airong Yang, Shuangxiu Wu, Yuntao Nie, Haifeng Shen, Jian Bai, Lin Wu, Fan Yang, Jun Wang. Spatio-temporal multiomics analysis reveals distinct molecular features in recurrent stage I non-small cell lung cancers after R0 tumor resection [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 618.