Prognosis Of Non-small Cell Lung Cancer Patients Research Articles

Prognostic role of targeted therapy in patients with multiple-site metastases from non-small- cell lung cancer

Aim: To evaluate the prognostic role of EGFR mutations in patients with multi-site metastases from non-small-cell lung cancer (NSCLC). Patients & methods: A total of 215 advanced NSCLC patients with multi-site metastases were included. The overall survival (OS) was the primary end point. Results: EGFR tyrosine kinase inhibitors (TKIs) significantly improved the OS in patients with brain metastases (p = 0.031) and bone metastases (p = 0.048). Meanwhile, it prolonged the OS in patients with lung or adrenal metastases, but not in patients with liver metastases. However, in patients without liver metastases, EGFR TKIs significantly improved the OS (p < 0.001). Conclusion: EGFR TKIs improved the prognosis in NSCLC patients with brain, bone, lung and adrenal metastases, but not in patients with liver metastases.

Correlations between circulating tumor cell phenotyping and 18F-fluorodeoxyglucose positron emission tomography uptake in non-small cell lung cancer.

The epithelial-to-mesenchymal transition (EMT) phenotype-based subsets of circulating tumor cells (CTCs) might be predictors of tumor progression. We evaluated the clinical properties of different phenotypic CTCs in patients with non-small cell lung cancer (NSCLC). Secondly, we explored the association between different phenotypic CTCs and the uptake of 18F-fluorodeoxyglucose (FDG) by the primary tumor on a positron emission tomographic (PET) scan. Venous blood samples from 34 pathologically confirmed Stage IIB-IVB NSCLC patients were collected prospectively. CTCs were immunoassayed using a SE-i·FISH®CTC kit. We identified CTCs into cytokeratin positive (CK+) and cytokeratin negative (CK-) phenotypes. CTC classifications were correlated with the maximum standardized uptake value (SUVmax) measured by 18F-fluorodeoxyglucose (FDG) positron emission tomography/computed tomography (PET/CT). Overall survival (OS) and progression-free survival (PFS) curves were produced using the Kaplan-Meier method. CTCs were detected in 91.2% of NSCLC patients. CTC counting was associated with TNM stage (P = 0.014) and distant metastasis (P = 0.007). The number of CK-CTCs was also positively associated with TNM stage (P = 0.022) and distant metastasis (P = 0.007). Both total CTC counting and CK-CTC counting did not show association with SUVmax value (P = 0.959, P = 0.903). Kaplan-Meier survival analysis demonstrated that patients with ≥ 7 CTCs had shorter OS (P = 0.003) and PFS (P = 0.001)relativeto patients with < 7 CTCs). Notably, the number of CK-CTCs can act as independent risk factors for PFS (P = 0.044) and OS (P = 0.043) in NSCLC patients. However, SUVmax value was not associated with OS (P = 0.895) and PFS (P = 0.686). The CTC subpopulations could be useful evidence for testing metastasis and prognosis in NSCLC patients. The SUVmax value of the primary tumor was not related to prognosis in patients with NSCLC.

Serum miR-1228-3p and miR-181a-5p as Noninvasive Biomarkers for Non-Small Cell Lung Cancer Diagnosis and Prognosis.

Background Lung cancer is the leading cause of cancer-related mortality worldwide, and non-small cell lung cancer (NSCLC) accounts for over 80% of all lung cancers. Serum microRNAs (miRNAs), due to their high stability, have the potential to become valuable noninvasive biomarkers. This present study was aimed to identify the serum miRNAs expression signatures for the diagnosis and prognosis of NSCLC using bioinformatics analysis. Methods A total of 12 miRNAs profiling studies have been identified in Pubmed, Gene Expression Omnibus (GEO), and ArreyExpress databases. Differentially expressed miRNAs (DEmiRNAs) were analyzed according to GEO2R online tool and RRA method from R. Then, prediction of DEmiRNAs' target genes from TargetScan, PicTar, miRDB, Tarbase, and miRanda database. Furthermore, we using reverse transcription– quantitative polymerase chain reaction (RT-qPCR) to evaluate the expression levels of DEmiRNAs in serum samples obtained from NSCLC patients and healthy controls. Subsequently, the clinical significance of the tested miRNAs was determined using receiver operating characteristic (ROC) analysis and Cox regression analysis. Results A total of 27 DEmiRNAs were identified and 5 of them (miR-1228-3p, miR-1228-5p, miR-133a-3p, miR-1273f, miR-545-3p) were significantly up-regulated and 4 of them (miR-181a-5p, miR-266-5p, miR-361-5p, miR-130a-3p) were significantly down-regulated in NSCLC patients compared with healthy controls. RT-qPCR validated that miR-1228-3p (P =0.006) and miR-181a-5p (P =0.030) were significantly differentially expressed in the serum of NSCLC patients and healthy controls. ROC analysis on miR-1228-3p and miR-181a-5p revealed the area under the curve (AUC) of 0.685 (95% confidence interval [CI], 0.563–0.806; P =0.006) and 0.647 (95% CI, 0.506–0.758; P =0.049). ROC analysis on miR-1228-3p combined miR-181a-5p revealed the AUC of 0.711 (95% CI, 0.593–0.828; P =0.002). Multivariate Cox regression analysis demonstrated that the high serum miR-1228-3p level was an independent factor for the poor prognosis of NSCLC patients. Conclusions Serum miR-1228-3p and miR-181a-5p are potential noninvasive biomarkers for the diagnosis and prognosis of NSCLC patients.

Katanin P60: a potential biomarker for lymph node metastasis and prognosis for non-small cell lung cancer

BackgroundThis study aimed to assess the correlation of katanin P60 expression with clinical characteristics and survival profiles of surgical non-small cell lung cancer (NSCLC) patients.MethodsTwo hundred and sixty-five primary NSCLC patients treated by surgical resection were retrospectively viewed. The expression of katanin P60 in the tumor specimen was detected by the immunohistochemical (IHC) staining assay. Preoperative clinical data were collected from patients’ medical records, and survival data were extracted from follow-up records.ResultsThere were 127 (47.9%) and 138 (52.1%) patients with katanin P60-low expression and -high expression, respectively; in addition, patients presenting katanin P60-high+, -high++, and -high+++ expression were 62 (23.4%), 63 (23.8%), and 13 (4.9%), respectively. Katanin P60 expression was correlated with lymph node (LYN) metastasis and advanced TNM stage but not pathological grade, tumor size, carcinoembryonic antigen (CEA) level or other non-tumor features in NSCLC patients. Regarding survival profiles, disease-free survival (DFS) and overall survival (OS) were both the lowest in katanin P60-high+++ expression patients, followed with katanin P60-high++ patients, katanin P60-high+ patients, and the highest in katanin P60-low expression patients. Further analysis illustrated that katanin P60-high expression was an independent predictive factor for unfavorable DFS and OS in NSCLC patients.ConclusionsKatanin P60 presents potential as a biomarker for lymph node metastasis and prognosis in NSCLC patients.

Circular RNA MYLK Promotes Glycolysis and Proliferation of Non-Small Cell Lung Cancer Cells by Sponging miR-195-5p and Increasing Glucose Transporter Member 3 Expression.

IntroductionCircular RNAs (circRNAs) are deregulated in many types of human cancers, including non-small cell lung cancer (NSCLC). In this study, we aimed to explore the functional role of circMYLK in NSCLC.Materials and MethodsThe expression levels of circMYLK and miR-195-5p in NSCLC tissues and cell lines were detected by RT-qPCR analysis. MTT assay, colony formation assay and transwell assay were performed to investigate the effects of circMYLK and miR-195-5p on the malignant phenotypes of NSCLC cells. The glucose consumption and lactate production of NSCLC cells were detected using commercial kits. The direct binding relation between circMYLK and miR-195-5p in NSCLC was predicted by bioinformatics analysis and validated by dual-luciferase reporter assay.ResultsThe results showed that circMYLK was significantly up-regulated in NSCLC tissues and cell lines, and its high expression was closely associated with deleterious clinicopathological characteristics and poor prognosis of NSCLC patients. Knockdown of circMYLK remarkably inhibited the malignant phenotypes of NSCLC cells, including proliferation, migration, invasion, glucose consumption and lactate production. Moreover, circMYLK was identified as a molecule sponge for miR-195-5p, and glucose transporter member 3 (GLUT3) was shown to be a target gene of miR-195-5p in NSCLC. Further rescue experiments revealed that the oncogenic effects of circMYLK on NSCLC cells could be largely abrogated by co-transfection with miR-195-5p mimic.ConclusionIn summary, our study provides convincing evidence that circMYLK serves as a tumor promoter in NSCLC and can be used as a potential therapeutic target for NSCLC patients.

Circular RNA circ_0000284 plays an oncogenic role in the progression of non-small cell lung cancer through the miR-377-3p-mediated PD-L1 promotion

BackgroundCircular RNAs (circRNAs), a subgroup of non-coding RNAs, are recognized as pivotal mediators in various types of cancers. CircRNA_0000284 (circ_0000284) was manifested to participate in the development of non-small cell lung cancer (NSCLC). The novel functional mechanism of circ_0000284 in NSCLC was investigated in our current study.MethodsWe exploited quantitative real-time polymerase chain reaction (qRT-PCR) to analyze the relative RNA (circRNA, miRNA and mRNA) expression. The assessment of cell proliferation and colony formation was executed by Cell Counting Kit-8 (CCK-8) and colony formation assay, respectively. Transwell assay was implemented to examine cell migration and invasion. All protein levels were assayed using western blot. The role of circ_0000284 in vivo was evaluated via xenograft model. The target relation was estimated by dual-luciferase reporter and RNA immunoprecipitation (RIP) assays.ResultsAs for the biological characterization, circ_0000284 was highly stable and localized in the cytoplasm. Circ_0000284 was up-regulated in NSCLC and could predict poor prognosis of NSCLC patients. Both in vitro and in vivo, down-regulation of circ_0000284 refrained tumorigenesis of NSCLC. Besides, microRNA-377-3p (miR-377-3p) was a miRNA target of circ_0000284, and targeted programmed death-ligand 1 (PD-L1). Circ_0000284 was a cancer-promoting circRNA in NSCLC via regulating the miR-377-3p/PD-L1 axis.ConclusionThus, our results unraveled that circ_0000284 facilitated the progression of NSCLC by up-regulating the PD-L1 expression as a competing endogenous RNA (ceRNA) of miR-377, possibly developing a different perspective in understanding the molecular pathogenesis of NSCLC.

MicroRNA-147 targets BDNF to inhibit cell proliferation, migration and invasion in non-small cell lung cancer.

Lung cancer is one of the most common cancers that threaten human life and health. Recently, microRNAs (miRNAs) have been shown to play a unique role in many malignancies. Although the dysregulation of miR-147 has been detected in non-small cell lung cancer (NSCLC), the biological function of miR-147 is still unknown in NSCLC. The expression of miR-147 was observed by real-time quantitative polymerase chain reaction (RT-qPCR). Methyl thiazolyl tetrazolium (MTT) and Transwell assays were used to investigate the function of miR-147 in NSCLC. Target genes of miR-147 were verified using dual luciferase reporter assay. Western blot analysis was used to explore the PI3K/AKT pathway. The expression of miR-147 was decreased in NSCLC tissues, which was associated with poor prognosis in NSCLC patients. Furthermore, overexpression of miR-147 inhibited the viability and metastasis of NSCLC cells. In addition, miR-147 inhibited epithelial-mesenchymal transition (EMT) and inactivated the PI3K/AKT pathway in NSCLC. Furthermore, miR-147 directly targets brain-derived neurotrophic factor (BDNF) and negatively regulates BDNF expression in NSCLC. Upregulation of BDNF attenuated the inhibitory effect of miR-147 in NSCLC. In conclusion, miR-147 inhibits cell proliferation, migration and invasion in NSCLC through suppressing BDNF expression.

Effect of uncommon and insensitive mutation on EGFR-TKI in treatment of non-small cell lung cancer.

e21652 Background: EGFR-TKI application is directed by the situation of mutations of epidermal growth factor receptor, in addition, the mutations of ALK, ROS-1,C-met, BRAF, Her-2, RET,NTRK1, PI3K, MEKI have potential effects on guiding treatment. This study aimed to investigate the uncommon and insensitive mutations and their effect on prognosis in non-small cell lung cancer (NSCLC) patients treated with EGFR-TKI. Methods: 21 NSCLC patients treated with EGFR-TKIs were enrolled, 9 with Gefitinib, 5 with Osimertinib and 7 with Icotinib. All patients were detected with targeted NGS 1000 gene panel. Afterwards, the EGFR sensitive mutation, uncommon mutation and mutations accompanied EGFR T790M were analyzed. Furthermore, the correlations of gene mutation with metastasis, OS and PFS were analyzed. SPSS 21.0 was used for statistical analysis, t test for continuous variables and χ2 test for classified variables. Results: In all the patients, EGFR p.[T790M] and p.[L858R] mutations were detected in 4.76% and 23.81% of the patients, respectively. In the 5 patients treated with Osimertinib, one had EGFR p.[T790M] mutation and curative effects was stable disease (SD). The mutations accompanied EGFR T790M included SMARCA4 p.[K1390Q] (2.23%), DNMT3A p.[F755S] (2.17%), MTOR p.[Y1450*] (2.15%), TPMT p.[L182R] (1.84%), etc. For the other four patients treated with Osimertinib, two of them were partial response (PR) and two were SD, accompanied with mutations such as EGFR p.[L858R] (55.00%), ARID1A p.[P16del] (2.83%), TP53 p.[R248W] (21.73%) and AR p.[Q60L] (4.24%). In addition, among all the mutations, 18 uncommon mutations were detected, in which ATM (23.26%), AR (23.26%) and MTOR (37.21%) were markedly associated with OS. Besides, the mutation frequency of 67 genes, such as ABCG2, AURKA, EPHA3 and ATR were correlated with OS. The mutation frequency of FGFR1 was significantly different between the patients with and without metastasis. The mutations of 22 genes, for instance ABCG2 p.[P21L], FAT1 p.[L398F], GNAS p.[F226V] and KMT2C p.[N2842Ilefs], were correlated with PFS during the treatment. Conclusions: EGFR-TKI has similar curative effects in EGFR p.[T790M] negative patients. Gene mutations, including uncommon mutations and EGFR insensitive mutations, can significantly affect the prognosis in NSCLC patients treated with EGFR-TKIs.

Effect of Cerebral Infarction on the Risk of Synchronous Brain Metastasis: Analysis of 307 Consecutive Patients of Newly Diagnosed Non-Small Cell Lung Cancer

Background: Brain metastasis (BM) is a common complication of patients with non-small cell lung cancer (NSCLC) and associated with a poor prognosis. The study aimed to evaluate the effect of cerebral infarction (CI) on the risk of BM in NSCLC for preventive therapy strategy. Methods: 307 patients with newly diagnosed NSCLC in Zhongda Hospital, Medical School of Southeast University from July 2013 to July 2018 were retrospectively analyzed. Depending on magnetic resonance imaging (MRI), the patients were divided into the BM group and the control group (without BM). Then, the prevalence of CI and baseline clinicopathological parameters were evaluated and compared between the two groups. Results: Out of the 307 patients, 204 patients (66.4%) had CI, and 52 patients (16.9%) had BM. Especially, the prevalence of CI in the NSCLC patients with BM was 84.6%, which was significantly higher than that of 62.7% in the NSCLC patients without BM (p = 0.002). Following univariate logistic regression analysis and the multivariate model, the results demonstrated that CI was a significant independent risk factor for BM in NSCLC (odds rate [OR], 2.921; 95% confidence interval [CI], 1.242-6.873; p = 0.014). Moreover, CI contributed to a worse prognosis in NSCLC patients with BM. Finally, dynamical trace confirmed CI could promote BM in NSCLC patients. Conclusions: CI could be associated with a metastatic tropism to the brain and then with an increased risk of BM in NSCLC patients. Therefore, the targeted intervention of the metastatic niche of CI could offer a promising approach for the prevention, prognostic evaluation, and therapy of BM in NSCLC patients for better clinical outcomes.

MicroRNA-374b mediates the initiation of non-small cell lung cancer by regulating ITGB1 and p53 expressions.

BackgroundPrevious studies have shown that microRNAs (miRNAs) play important roles in the pathogenesis of human cancers. This study aims to clarify the role of miR‐374b in non‐small cell lung cancer (NSCLC).MethodsIn this study, RT‐qPCR and western blot analysis were used to measure mRNA and protein expression. The regulatory mechanism of miR‐374b/ITGB1 was investigated by dual‐luciferase reporter, CCK‐8, and transwell assays.ResultsMiR‐374b expression was reduced in NSCLC tissues and associated with lymph node metastasis, tumor stage and prognosis in NSCLC patients. Functionally, overexpression of miR‐374b inhibited cell viability and metastasis in NSCLC. In addition, miR‐374b blocked EMT and promoted p53 expression in NSCLC. MiR‐374b was found to directly target ITGB1. Furthermore, upregulation of ITGB1 weakened the antitumor effect of miR‐374b in NSCLC.ConclusionsMiR‐374b inhibits the tumorigenesis of NSCLC by downregulating ITGB1 and upregulating p53.

A Shallow Convolutional Neural Network Predicts Prognosis of Lung Cancer Patients in Multi-Institutional CT-Image Data.

Lung cancer is the most common fatal malignancy in adults worldwide, and non-small cell lung cancer (NSCLC) accounts for 85% of lung cancer diagnoses. Computed tomography (CT) is routinely used in clinical practice to determine lung cancer treatment and assess prognosis. Here, we developed LungNet, a shallow convolutional neural network for predicting outcomes of NSCLC patients. We trained and evaluated LungNet on four independent cohorts of NSCLC patients from four medical centers: Stanford Hospital (n = 129), H. Lee Moffitt Cancer Center and Research Institute (n = 185), MAASTRO Clinic (n = 311) and Charité – Universitätsmedizin (n=84). We show that outcomes from LungNet are predictive of overall survival in all four independent survival cohorts as measured by concordance indices of 0.62, 0.62, 0.62 and 0.58 on cohorts 1, 2, 3, and 4, respectively. Further, the survival model can be used, via transfer learning, for classifying benign vs malignant nodules on the Lung Image Database Consortium (n = 1010), with improved performance (AUC=0.85) versus training from scratch (AUC=0.82). LungNet can be used as a noninvasive predictor for prognosis in NSCLC patients and can facilitate interpretation of CT images for lung cancer stratification and prognostication.

Expression of Circulating Microparticles for the Diagnosis of Non-small Cell Lung Cancer: Clinicopathological Correlations and Prognostic Value

Increased values of circulating microparticles (MPs) have been reported in solid tumors including non-small cell lung cancer (NSCLC). We therefore investigated the utility of baseline MPs in the clinical setting of patients with NSCLC. Quantification of MPs in the plasma was performed by flowcytometry. Baseline MP values were correlated with clinical patients' characteristics, estimated tumor volume (ETV) and treatment response. Receiver operating characteristics (ROC) curves were plotted to discriminate between patients and controls in order to determine the diagnostic value of circulating MPs in NSCLC. Our prospective study included 134 NSCLC patients (98 at initial diagnosis, ID and 36 at relapse, R) and 30 healthy individuals. The mean of baseline MP numbers was significantly higher in patients presented either at ID or R than in controls (p<0.0001). Basal MP numbers were inversely correlated with ETV values (p=0.04). In addition, the difference in MP levels at diagnosis was significant according to tumor histology (p=0.02) and primary tumor size (p=0.0007). Using ROC analysis, the optimal cutoff value for baseline MPs was 1307 events/µL with a sensitivity and a specificity of 67.3% and 90.0%, respectively. High MPs expression was significantly associated with low-level smoking degree (p=0.001), non-squamous cell types (p=0.017) and decreased tumor size (p=0.003). Our results suggest that high baseline MP values could be an indicator of tumor growth inhibition in NSSLC. Furthermore, high expression of circulating MPs at diagnosis might predict good prognosis in NSCLC patients.

ATM gene polymorphisms are associated with poor prognosis of non-small cell lung cancer receiving radiation therapy.

We investigated the prognostic significance of ataxia telangiectasia mutated (ATM) single nucleotide polymorphisms (SNPs) in 720 Han Chinese non-small cell lung cancer (NSCLC) patients who underwent radiation or chemoradiation therapy. Kaplan-Meier survival curves showed that overall survival (OS) and disease-free survival (DFS) rates were significantly associated with two ATM SNPs, rs664143 and rs189037. Patients with the rs664143 GA or AA genotype had poorer DFS (hazard ratio (HR) = 1.40, 95% confidence interval (CI) = 1.05–1.86, P = 0.021) and OS (HR = 1.28, 95%CI = 1.12–1.78, P = 0.040) than those with the rs664143 GG phenotype. Patients with the rs189037 AG/GG genotypes had poorer prognoses than those with the rs189037 AA genotype (AG/GG vs. AA: DFS, HR = 1.44, 95%CI = 1.06–1.95, P=0.019; OS, HR = 1.16, 95%CI = 1.16–1.17–2.21, P=0.004). These results were confirmed by subgroup analysis based on clinical factors such as smoking, histology, tumor stage, treatment, and radiation dose, all of which were significantly associated with DFS and OS rates in NSCLC patients. These findings show that ATM rs664143 and rs189037 variants determine prognosis in NSCLC patients that have undergone radiation or chemoradiation therapies.

MicroRNA-597 inhibits NSCLC progression through negatively regulating CDK2 expression.

Previous studies have shown that microRNA-597 serves as a tumor suppressor gene. However, the role of microRNA-597 in non-small cell lung cancer (NSCLC) has not been fully elucidated. Therefore, the aim of this study was to investigate the expression of microRNA-597 in NSCLC, and to further explore the possible underlying mechanism. Real-time quantitative polymerase chain reaction (qPCR) was performed to examine microRNA-597 level in tumor tissues and para-cancerous normal tissues collected from 50 patients with NSCLC. The interplay between microRNA-597 expression and clinical indicators, as well as prognosis of NSCLC patients, was analyzed. Meanwhile, qPCR was used to verify microRNA-59 level in NSCLC cell lines. Subsequently, microRNA-597 overexpression and knockdown models were constructed using lentivirus in NSCLC cell lines (including H1299 and PC-9). The impacts of microRNA-597 on the biological functions of NSCLC cells were evaluated using cell counting kit-8 (CCK-8), colony formation, and 5-Ethynyl-2'-deoxyuridine (EdU) assay, respectively. Finally, luciferase reporter gene assay and recovery experiment were performed to investigate the underlying molecular mechanism. QPCR results indicated that microRNA-597 level in NSCLC tissues was remarkably lower than that of adjacent normal tissues, and the difference was statistically significant (p<0.05). Compared with patients with high expression of microRNA-597, patients with low expression of microRNA-597 exhibited significantly higher incidence of pathological stage and lower overall survival rate (p<0.05). Similarly, compared with NC group, the proliferation ability of NSCLC cells was remarkably weakened in microRNA-597 overexpression group (p<0.05). However, the opposite results were observed in microRNA-597 inhibitor group (p<0.05). CDK2 expression was found remarkably elevated in NSCLC cell lines as well as in tissue samples CDK2 expression. Meanwhile, CDK2 expression was negatively correlated with microRNA-597 expression. Luciferase reporter gene assay demonstrated that overexpression of CDK2 could significantly attenuate the luciferase activity of wild-type microRNA-597 vector without attenuating that of mutant vector CDK2 expression. This further suggested that microRNA-597 could target bind to CDK2. Furthermore, cell recovery experiment revealed that CDK2 could reverse the impact of microRNA-597 on the malignant progression of NSCLC. MicroRNA-597 expression was significantly down-regulated in NSCLC tissues, as well as cell lines. Meanwhile, microRNA-597 expression was associated with the pathological staging and poor prognosis of patients with NSCLC. In addition, microRNA-597 might suppress the malignant progression of NSCLC through the regulation of CDK2.

Up-Regulation of FSTL3, Regulated by lncRNA DSCAM-AS1/miR-122-5p Axis, Promotes Proliferation and Migration of Non-Small Cell Lung Cancer Cells.

BackgroundFollistatin-like 3 (FSTL3) binds and inactivates activin, a growth factor with cell growth and differentiation. Previous studies reported that it is overexpressed in invasive breast cancers, and its expression and function in non-small cell lung cancer (NSCLC) remain unclear.Materials and MethodsImmunohistochemistry was employed to probe the expression of FSTL3 in NSCLC tissues. Real-time PCR (RT-PCR) was applied to detect the expression of lncRNA DSCAM-AS1 and miR-122-5p. A549 cells and H1299 cells were used as cell models. The biological influence of FSTL3 on cells was studied using CCK-8 assay, wound healing assay and transwell assay in vitro, respectively. In vivo subcutaneous xenotransplanted tumor model and tail vein injection model in mice were also constructed to validate the roles of FSTL3. Interactions between miR-122-5p and FSTL3, DSCAM-AS1 and miR-122-5p were determined by bioinformatics analysis, RT-PCR, and dual-luciferase reporter assay.ResultsFSTL3 and DSCAM-AS1 were remarkably up-regulated in NSCLC samples, and miR-122-5p was down-regulated. FSTL3 was associated with worse prognosis of NSCLC patients. FSTL3 knockdown markedly inhibited the viability, migration and invasion of NSCLCs in vitro and in vivo. DSCAM-AS1 could down-regulate miR-122-5p via sponging it, and FSTL3 was a target gene of miR-122-5p.ConclusionTaken together, our study identified that FSTL3 was a new oncogene of NSCLC, which was regulated by DSCAM-AS1 and miR-122-5p. These findings suggested that FSTL3, DSCAM-AS1 and miR-122-5p might serve as a new valuable therapeutic target for NSCLC.

CircFOXM1 promotes proliferation of non-small cell lung carcinoma cells by acting as a ceRNA to upregulate FAM83D

BackgroundBiological role and clinical significance of circular RNAs (circRNAs) remain largely unknown. Herein, we aimed to investigate biological function, molecular mechanism, and clinical significance of a circular RNA FOXM1 (circFOXM1) in non-small cell lung cancer (NSCLC).MethodsExpression of circFOXM1 was measured in 48 paired samples of NSCLC by qRT-PCR. Functional roles of circFOXM1 on tumor cells were explored by in vitro and in vivo assays. Transcriptome sequencing was employed to screen the molecules involved in circFOXM1 regulatory network. RNA immunoprecipitation, luciferase analysis, RNA pull-down, and rescue assay were used to investigate potential mechanisms of circFOXM1.ResultsWe found that circFOXM1 was significantly upregulated in NSCLC tissues, and its upregulation was positively correlated with advanced clinical stage and poor prognosis of NSCLC patients. Gain or loss-of-function assay showed that circFOXM1 promoted cell proliferation and cell cycle progression. In vivo assays showed that silencing circFOXM1 inhibited xenograft tumor growth. Mechanically, transcriptome sequencing data indicated that silencing circFOXM1 led to the downregulation of cell cycle-related mRNAs. RNA pull-down and dual-luciferase reporter assay suggested that circFOXM1 could bind to miR-614, and FAM83D was an essential gene involved in the circFOXM1/miR-614 regulatory network.ConclusionscircFOXM1promotes NSCLC progression by interacting with miR-614 and thus inactivating the function of miR-614, which will further release the suppression of FAM83D. circFOXM1/miR-614/FAM83D regulatory network may serve as a potential therapeutic target for NSCLC patients.

MiR-519d targets HER3 and can be used as a potential serum biomarker for non-small cell lung cancer.

Development of specific serum biomarkers is essential to improve diagnosis and prognosis of non-small cell lung cancer (NSCLC). Here, we show that serum and tissue levels of miR-519d are significantly decreased in NSCLC patients. The low expression of miR-519d is associated with lymph node metastases, clinical stage, and a poor prognosis in NSCLC patients. In addition, ROC analysis demonstrated that the serum miR-519d levels can distinguish NSCLC patients from healthy controls. MiR-519d inhibits proliferation, migration, and invasion by lung cancer cells, indicating that it may function as a tumor suppressor in lung cancer. Furthermore, our data demonstrate that HER3 is a target gene of miR-519d in lung cancer cells, and show that by targeting HER3, miR-519d inhibits the PI3K/Akt pathway. These findings demonstrate that the miR-519d levels are decreased in serum and tumor tissues of NSCLC patients, and indicate that miR-519d regulates NSCLC progression by targeting HER3. MiR-519d could potentially serve as a novel serum biomarker for NSCLC.

Clinical course of non-small cell lung cancer patients with dry pleural dissemination: Retrospective observational study.

We investigated the prognosis of patients with dry pleural dissemination (DPD) of non-small cell lung cancer (NSCLC) and the risk factors of developing to malignant pleural effusion (MPE).We retrospectively reviewed 104 patients with NSCLC and DPD, confirmed surgically from 1996 to 2016. Incidence rate and risk factors of MPE were analyzed statistically. The prognosis of NSCLC patients with MPE was evaluated using the Kaplan–Meier method.The most common histologic type was adenocarcinoma in 95 (91.3%) patients. The median follow-up duration was 65.5 months and the median survival time was 37.7 months. MPE developed in 51 (49%) patients, and the median effusion-free interval was 41.9 months. The median survival time of the patients with and without MPE was not different (41.3 vs 31.7 months, P = .16). No predictive factors for the development of MPE were identified. Fifteen (14.4%) patients underwent invasive procedures for the management of MPE.Almost half of all patients with NSCLC and DPD experienced MPE, and 14.4% patients developed symptomatic MPE requiring invasive procedures. MPE in DPD did not affect the survival in NSCLC patients.

USP3 promotes proliferation of non-small cell lung cancer through regulating RBM4.

Previous studies have shown that ubiquitin specific protease 3 (USP3) is an oncogene. However, the role of USP3 in non-small cell lung cancer (NSCLC) has not been reported. This study aims to explore the expression characteristics of USP3 in NSCLC, and its regulation on the proliferative capacity of NSCLC cells. Quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) was performed to examine the expression levels of USP3 and RNA Binding Motif 4 (RBM4) in 42 pairs of tumor tissues and adjacent tissue specimens collected from NSCLC patients. Meanwhile, the correlation between the messenger ribonucleic acid (mRNA) expressions of USP3 and RBM4, and the clinical indicators and prognosis of NSCLC patients were analyzed. At the same time, mRNA expression of USP3 in NSCLC cell lines was further verified by the qRT-PCR method. In addition, USP3 knockdown and overexpression models were constructed using lentivirus in NSCLC cell lines H1299 and SPCA1. Cell counting kit-8 (CCK-8), cell colony formation, and 5-Ethynyl-2'-deoxyuridine (EDU) assays were performed to evaluate the influence of USP3 on proliferative capacity in NSCLC cells. Finally, Dual-Luciferase reporter assay and rescue experiments were conducted to further explore its underlying molecular mechanism. In this experiment, qRT-PCR results revealed that the expression level of USP3 in tumor tissues of NSCLC patients was remarkably higher than that in adjacent tissues, and the difference was statistically significant. Compared with NSCLC patients with low expression of USP3, those with high USP3 expression suffered much more advanced pathology stage and lower overall survival rate. Proliferation ability of NSCLC cells overexpressing USP3 was remarkably enhanced, while the opposite result was observed in the USP3 knockdown group. Subsequently, RBM4 expression in NSCLC tissue samples was found to be significantly reduced and negatively correlated with USP3 level. In addition, the result of Dual-Luciferase reporter assay demonstrated that USP3 can be targeted by RBM4. Rescue experiments revealed that RBM4 was responsible for NSCLC progression regulated by USP3. The above studies indicated that USP3 expression was remarkably up-regulated in NSCLC tissues, which was closely related to the pathological staging and poor prognosis of NSCLC patients. Therefore, USP3 might accelerate the proliferation of NSCLC cells via regulating RBM4.

Prognostic value of cripto-1 expression in non-small-cell lung cancer patients: a systematic review and meta-analysis.

Aim: This systematic review and meta-analysis aimed to analyze the association between cripto-1 expression and prognosis as well as clinicopathological features of non-small-cell lung cancer (NSCLC) patients. Methods: The electronic databases for all articles about NSCLC and cripto-1 expression were searched. Results: Twelve articles were enrolled in this meta-analysis (3130 samples). In NSCLC patients, cripto-1 was expressed higher than in normal tissues. Cripto-1 expression was closely correlated with lymph node metastasis, histological differentiation and advanced clinical stage of NSCLC patients, but not related to smoking, age and gender. Pooled hazard ratios found that high cripto-1 expression had poor overall survival and progression-free survival. Conclusion: Cripto-1 could serve as a novel biomarker for predicting poor prognosis in NSCLC patients.