Value of Endobronchial Ultrasound-Guided Transbronchial Needle Aspiration to Diagnose the Lung Nodules Related Enlarged Mediastinal Lymph Nodes.

Background: To investigate the value of endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) in the diagnosis of lung nodules related enlarged mediastinal lymph nodes (MLNs). Methods: Clinical data of 108 patients with lung nodules related enlarged MLNs who underwent EBUS-TBNA in our single center were retrospectively analyzed from January 2020 to December 2021. The sensitivity and specificity of EBUS-TBNA in malignancy diagnosis were evaluated. Associations between ultrasonic image measurement indexes and malignancy diagnosis were explored. The receiver operating characteristic (ROC) curve of these indexes, the area under curve (AUC), and the corresponding cut-off values were calculated to predict malignant MLNs. Results: Sensitivity, specificity, and accuracy of EBUS-TBNA in the diagnosis of lung nodules related malignant MLNs were 89.47%, 100%, and 92.59%, respectively. There were significantly higher proportions of malignant MLNs with clear boundary, short diameter ≥1 cm, lower long to short diameter ratio, abundant flow of blood, and destructed medulla than that of benign ones (P < .05). According to ROC curve analysis, the cut-off value of short diameters for predicting malignant MLNs was 1.085 cm, and the AUC was 0.796 (95% confidence interval: 0.724-0.868, P < .001). Corresponding sensitivity and specificity were 61.36% and 80.00%, respectively. The cut-off value of the long to short diameter ratio for predicting malignancy was 1.405, and the AUC was 0.697 (95% confidence interval: 0.609-0.790, P < .001). Corresponding sensitivity and specificity were 70.00% and 71.97%, respectively. Conclusion: EBUS-TBNA has a satisfactory accuracy of lung nodules related MLNs diagnosis. Short diameters and long to short diameter ratio of lung nodules related MLNs in ultrasonic image may contribute to the prediction of malignant lymph nodes.

Circ-ATIC Serves as a Sponge of miR-326 to Accelerate Esophageal Squamous Cell Carcinoma Progression by Targeting ID1.

In the previous studies, circular RNA (circRNA) has been shown to be closely related to the occurrence and development of various cancers. However, the role and mechanism of circ-ATIC in the progression of esophageal squamous cell carcinoma (ESCC) is not yet clear. Quantitative real-time PCR was used to detect the expression levels of circ-ATIC, microRNA (miR)-326 and inhibitor of DNA binding 1 (ID1) in tissues (n = 50) and cells. Cell counting kit 8 assay, colony formation assay, flow cytometry, wound-healing assay and transwell assay were performed to measure the proliferation, apoptosis, migration, and invasion of cells. In addition, the oxidative stress of cells was evaluated by detecting the productions of superoxide dismutase and malondialdehyde. Animal studies were implied to explore the role of circ-ATIC in ESCC tumor growth. The relationship between circ-ATIC and miR-326 or ID1 was determined by dual-luciferase reporter assay and RNA immunoprecipitation assay. Additionally, the protein expression of ID1 was examined by western blot assay. Circ-ATIC was found to be upregulated in ESCC tissues and cells. Silenced circ-ATIC suppressed the proliferation, migration, invasion, promoted the apoptosis and oxidative stress of ESCC cells. The tumor growth of ESCC also was inhibited by circ-ATIC knockdown. Furthermore, we found that circ-ATIC could sponge miR-326, and miR-326 could target ID1. The rescue experiments revealed that miR-326 inhibitor could reverse the negative regulation of circ-ATIC silencing on ESCC progression, and ID1 overexpression also inverted the inhibitory effect of miR-326 on ESCC progression. In addition, we confirmed that the expression of ID1 was positively regulated by circ-ATIC. Our study showed that circ-ATIC facilitated the progression of ESCC by regulating the miR-326/ID1 axis, indicating that circ-ATIC might be a target for ESCC treatment.

MiR-519d-3p enhances the sensitivity of non-small-cell lung cancer to tyrosine kinase inhibitors.

Non-small cell lung cancer (NSCLC) is the leading cause of cancer mortality worldwide. Tyrosine kinase inhibitors (TKIs) are currently the most effective chemotherapy for NSCLC. However, most cancer patients develop TKI resistance at tumor relapse stage. We firstly measured the expression change of miR-519d-3p in TKI resistance NSCLC cells. We then ectopically expressed miR-519-3p in TKI resistant cells to study its functional impact on cell proliferation, migration, invasion and cell sensitivity to gefitinib. The downstream target of miR-519-3p was identified by bioinformatics and validated in luciferase reporter assay and western blotting analysis. We also studied the reversing effect of the candidate target in NSCLC cells expressing miR-519d-3p. Lastly, we compared the miR-519d-3p level in NSCLC patients with good or poor response to gefitinib. miR-519d-3p level was downregulated in TKI resistant NSCLC cells. The restoration of miR-519d-3p in these NSCLC cells inhibited cell proliferation, invasion and migration; enhanced cell sensitivity to gefitinib. EPAS1 was identified and validated as downstream target of miR-519d-3p. Co-expressing EPAS1 antagonized the inhibitory effect of miR-519d-3p on NSCLC cells. MiR-519d-3p was downregulated in NSCLC patients with poor response to gefitinib. Targeting miR-519d-3p/EPAS1 axis may provide alternative treatment for TKI-resistant NSCLC.

EPAS1 promotes peritoneal carcinomatosis of non-small-cell lung cancer by enhancing mesothelial-mesenchymal transition.

BACKGROUND: Non-small-cell lung cancer (NSCLC) is a major cause of cancer-related death globally. Endothelial PAS domain-containing protein 1 (EPAS1) is a homolog of the hypoxia-inducible factor 1α and has been reported to confer tyrosine kinase inhibitor (TKI) resistance in NSCLC, but its role in peritoneal carcinomatosis of NSCLC is unknown. METHODS: PC14HM, a high metastatic potential subline of NSCLC cell line PC14, was derived. Stable shRNA knockdown of EPAS1 was then established in PC14HM cells and subjected to assessment regarding the effects on proliferation and viability, xenograft tumor growth, metastatic potential, mesothelial-mesenchymal transition (MMT)-related characteristics and peritoneal carcinomatosis in a mouse model. RESULTS: EPAS1 expression was elevated in PC14HM cells. Knockdown of EPAS1 inhibited the proliferation and viability of PC14HM cells in vitro and suppressed tumorigenesis in vivo. In addition, the metastatic features and in vitro productions of MMT-inducing factors in PC14HM cells was also associated with EPAS1. More importantly, knockdown of EPAS1 drastically suppressed peritoneal carcinomatosis of PC14HM cells in vivo. CONCLUSION: EPAS1 promotes peritoneal carcinomatosis of NSCLC through enhancement of MMT and could therefore serve as a prognostic marker or a therapeutic target in treating NSCLC, particularly in patients with peritoneal carcinomatosis.

Long Non-coding RNA MIAT Mediates Non-small Cell Lung Cancer Development Through Regulating the miR-128-3p/PELI3 Axis.

In this study, we set out to characterize the expression status of long non-coding RNA (lncRNA) Myocardial Infarction Associated Transcript (MIAT) in non-small cell lung cancer (NSCLC) and elucidate its mechanistic contribution to this disease. Relative expression levels of MIAT, Pellino E3 Ubiquitin Protein Ligase Family Member 3 (PELI3), and microRNA (miR)-128-3p were analyzed by real-time polymerase chain reaction. PELI3 protein level was determined by immunoblotting. Cell viability and proliferation were evaluated by the MTT assay and colony formation assay, respectively. Cell invasion and migration were assessed by wound-healing closure and transwell assays, respectively. The regulatory actions of miR-128-3p on both MIAT and PELI3 were interrogated by luciferase reporter assay. We demonstrated the aberrant upregulation of MIAT in NSCLC and its association with tumor progression. We further uncovered the negative correlation among MIAT, PELI3, and miR-128-3p. MIAT deficiency significantly compromised cell viability, proliferation, invasion, and migration, while increased miR-128-3p and decreased PELI3 expressions. Application of miR-128-3p inhibitor significantly stimulated luciferase activities driven by both MIAT and PELI3 promoter and phenotypically promoted cell viability, proliferation, migration, and invasion. Our study highlighted the mechanistic contribution of the MIAT/miR-128-3p/PELI3 signaling cascade in NSCLC.

LncRNA DANCR Promotes Lung Cancer by Sequestering miR-216a.

BACKGROUND: Long noncoding RNAs (lncRNAs) are a new class of cancer regulators. Here, we aimed to investigate the diagnostic and therapeutic values of an lncRNA, differentiation antagonizing noncoding RNA (DANCR), in lung cancer. METHODS: Real-time polymerase chain reaction was used to compare DANCR levels in normal and cancerous lung tissues as well as lung cancer cells. Lentiviral transduction was used to induce DANCR overexpression or silencing in vitro, followed by monitoring cell proliferation, colony formation, and changes in microRNA-216a (miR-216a) expression. DANCR-specific small hairpin RNA transduction was used to establish cells with stable DANCR knockdown, and silenced cells were used to initiate lung tumor xenografts, followed by monitoring tumor growth. RESULTS: DANCR upregulation was seen in lung cancer, particularly in high-grade lung cancer tissues and aggressive cancer cells. Ectopic DANCR expression induced lung cancer cell proliferation and colony formation, whereas DANCR silencing induced opposing effects. The miR-216a level in cancer cells was negatively correlated with DANCR expression. The DANCR knockdown reduced the growth of tumor xenografts in vivo. CONCLUSION: DANCR upregulation is a potential indicator of aggressive lung cancer. Silencing of DANCR has great potential as a potent therapeutic strategy in lung cancer.

LncRNA PCAT-1 promotes tumour growth and chemoresistance of oesophageal cancer to cisplatin.

Oesophageal cancer (OC) is one of the most fatal malignancies in the world, and chemoresistance restricts the therapeutic outcome of OC. Long noncoding RNA (lncRNA) was reported to play roles in multiple cancer types. Yet, the function of lncRNA in chemoresistance of OC has not been reported. A lncRNA gene, PCAT-1, showed higher expression in OC tissues, especially higher in secondary OC compared with normal mucosa tissues. Overexpression of PCAT-1 increased the proliferation rate and growth of OC cells. Inhibition of PCAT-1 decreased proliferation and growth of OC cells, and increased cisplatin chemosensitivity. In a mouse OC xenograft model, PCAT-1 inhibition repressed OC growth in vivo. Therefore, PCAT-1 may potentially serve as a therapeutic target for treating OC. PCAT-1 promotes development of OC and represses the chemoresistance of OC to cisplatin, and silencing of PCAT-1 may be a therapeutic strategy for treating OC.

MicroRNA-124 suppresses proliferation and glycolysis in non-small cell lung cancer cells by targeting AKT-GLUT1/HKII.

Non-small cell lung cancer accounts for 85% of all types of lung cancer and is the leading cause of worldwide cancer-associated mortalities. MiR-124 is epigenetically silenced in various types of cancer and plays important roles in tumor development and progression. MiR-124 was also significantly downregulated in non-small cell lung cancer patients. Glycolysis has been considered as a feature of cancer cells; hypoxia-inducible factor 1-alpha/beta and Akt are key enzymes in the regulation of glycolysis and energy metabolism in cancer cells. However, the role of miR-124 in non-small cell lung cancer cell proliferation, glycolysis, and energy metabolism remains unknown. In this research, cell proliferation was investigated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; furthermore, glucose consumption and lactic acid production were assessed; adenosine triphosphate content and NAD+/NADH were also detected. These tests were conducted using the normal non-small cell lung cancer cell line A549, which was transfected variedly with miR-mimics, miR-124 mimics, miR-124 inhibitor, pc-DNA3.1(+)-AKT1, and pc-DNA3.1(+)-AKT2 plasmid. Here, we show that miR-124 overexpression directly decreased cell growth, glucose consumption, lactate production, and energy metabolism. MiR-124 also negatively regulates glycolysis rate-limiting enzymes, glucose transporter 1 and hexokinase II. Our results also showed that miR-124 negatively regulates AKT1 and AKT2 but no regulatory effect on hypoxia-inducible factor 1-alpha/beta. Overexpression of AKT reverses the inhibitory effect of miR-124 on cell proliferation and glycolytic metabolism in non-small cell lung cancer. AKT inhibition blocks miR-124 silencing-induced AKT1/2, glucose transporter 1, hexokinase II activation, cell proliferation, and glycolytic or energy metabolism changes. In summary, this study demonstrated that miR-124 is able to inhibit proliferation, glycolysis, and energy metabolism, potentially by targeting AKT1/2-glucose transporter 1/hexokinase II in non-small cell lung cancer cells.

Improving the Accuracy of Daily PM2.5 Distributions Derived from the Fusion of Ground-Level Measurements with Aerosol Optical Depth Observations, a Case Study in North China.

The accuracy in estimated fine particulate matter concentrations (PM2.5), obtained by fusing of station-based measurements and satellite-based aerosol optical depth (AOD), is often reduced without accounting for the spatial and temporal variations in PM2.5 and missing AOD observations. In this study, a city-specific linear regression model was first developed to fill in missing AOD data. A novel interpolation-based variable, PM2.5 spatial interpolator (PMSI2.5), was also introduced to account for the spatial dependence in PM2.5 across grid cells. A Bayesian hierarchical model was then developed to estimate spatiotemporal relationships between AOD and PM2.5. These methods were evaluated through a city-specific 10-fold cross-validation procedure in a case study in North China in 2014. The cross validation R(2) was 0.61 when PMSI2.5 was included and 0.48 when PMSI2.5 was excluded. The gap-filled AOD values also effectively improved predicted PM2.5 concentrations with an R(2) = 0.78. Daily ground-level PM2.5 concentration fields at a 12 km resolution were predicted with complete spatial and temporal coverage. This study also indicates that model prediction performance should be assessed by accounting for monitor clustering due to the potential misinterpretation of model accuracy in spatial prediction when validation monitors are randomly selected.

MicroRNA-200a Targets EGFR and c-Met to Inhibit Migration, Invasion, and Gefitinib Resistance in Non-Small Cell Lung Cancer.

Lung cancer, especially non-small cell lung cancer (NSCLC), is the major cause of cancer death worldwide. Mutations in epidermal growth factor receptor (EGFR) and hepatocyte growth factor receptor (c-Met), both of which are receptor tyrosine kinases, have been identified in a considerable percentage of NSCLC patients. EGFR and c-Met share the same downstream pathways and cooperate not only in promoting metastasis but also in conferring resistance to tyrosine kinase inhibitor (TKI) therapies in NSCLC. MicroRNAs (miRNAs) are a family of small non-coding RNAs, usually 21-25 nucleotides long, and are critical in regulating gene expression. Abnormal miRNA expression has been implicated in the initiation and progression in many forms of cancers, including lung cancer. In this study, we found that miR-200a is downregulated in NSCLC cells, where it directly targets the 3'-UTR of both EGFR and c-Met mRNA. Overexpression of miR-200a in NSCLC cells significantly downregulates both EGFR and c-Met levels and severely inhibits cell migration and invasion. Moreover, in NSCLC cell lines that are resistant to gefitinib, a drug often used in TKI therapies to treat NSCLC, miR-200a expression is able to render the cells much more sensitive to the drug treatment.

Daily Estimation of Ground-Level PM2.5 Concentrations over Beijing Using 3 km Resolution MODIS AOD.

Estimating exposures to PM2.5 within urban areas requires surface PM2.5 concentrations at high temporal and spatial resolutions. We developed a mixed effects model to derive daily estimations of surface PM2.5 levels in Beijing, using the 3 km resolution satellite aerosol optical depth (AOD) calibrated daily by the newly available high-density surface measurements. The mixed effects model accounts for daily variations of AOD-PM2.5 relationships and shows good performance in model predictions (R(2) of 0.81-0.83) and cross-validations (R(2) of 0.75-0.79). Satellite derived population-weighted mean PM2.5 for Beijing was 51.2 µg/m(3) over the study period (Mar 2013 to Apr 2014), 46% higher than China's annual-mean PM2.5 standard of 35 µg/m(3). We estimated that more than 19.2 million people (98% of Beijing's population) are exposed to harmful level of long-term PM2.5 pollution. During 25% of the days with model data, the population-weighted mean PM2.5 exceeded China's daily PM2.5 standard of 75 µg/m(3). Predicted high-resolution daily PM2.5 maps are useful to identify pollution "hot spots" and estimate short- and long-term exposure. We further demonstrated that a good calibration of the satellite data requires a relatively large number of ground-level PM2.5 monitoring sites and more are still needed in Beijing.

A novel minimally invasive fixation method for flail chest management in a Canine model: an animal research.

BACKGROUND: Multiple rib fractures can lead to flail chest with up to 35% mortality rate due to severe pulmonary complications. Current treatments of flail chest remain controversial. Studies have shown that surgical treatments can improve outcomes and reduce mortality, comparing to non-operative treatments. Current surgical fixation methods focus on stabilization of ribs on the outward facing side, and they require division of intercostal muscles. Damages to surrounding nerves and vessels may lead to chronic pain. This study tests a novel interior fixation method that minimizes neurovascular injuries. METHODS: Twelve healthy canines were divided in two surgical operation groups for exterior and interior fixation using titanium metal plates. Osteotomy with oblique fractures was prepared under general anesthesia. Exterior fixation was performed in open surgery. Interior fixation was minimally invasive using custom made tools including a flexible shaft extension screwdriver, solid plate stand, guiding wire loop and metal plates with threaded holes. RESULTS: Respiratory and cardiovascular functions (RR, PO2, PCO2, SpO2, and HR) together with body temperature were measured before anesthesia and within 48 h after surgery. The difference in measurements was not statistically significant between the two groups before surgery with P values greater than 0.05. However, the interior group canines had better RR and PO2 values starting from the 24th hour, and better PCO2, SpO2, and HR values starting from the 48th hour. It took longer operation time to complete the minimally invasive interior fixation surgery (P value less than 0.001), but the total blood loss was less than the exterior fixation group (P value less than 0.001). Results also showed that interior group canines suffered less pain, and they had quicker recovery in gastrointestinal and physical mobility. CONCLUSIONS: The investigative interior fixation method was safe and effective in rib stabilization on a canine rib fracture model, comparing to the exterior fixation method. The interior fixation was minimally invasive, with less damages to tissues and nerves surrounding the ribs, leading to better postoperative outcomes.

Curcumin increases crizotinib sensitivity through the inactivation of autophagy via epigenetic modulation of the miR-142-5p/Ulk1 axis in non-small cell lung cancer.

Drug resistance is a critical factor responsible for the recurrence of non-small cell lung cancer (NSCLC). Previous studies suggest that curcumin acts as a chemosensitizer and radiosensitizer in human malignancies, but the underlying mechanism remains elusive. In the present study, we explored how curcumin regulates the expression of miR-142-5p and sensitizes NSCLC cells to crizotinib. We found that miR-142-5p is significantly downregulated in NSCLC tissue samples and cell lines. Curcumin could increase crizotinib cytotoxicity by epigenetically restoring the expression of miR-142-5p. Furthermore, curcumin treatment suppressed the expression of DNA methylation-related enzymes, including DNMT1, DNMT3A, and DNMT3B, in NSCLC cells. In addition, the upregulation of miR-142-5p expression increased crizotinib cytotoxicity and induced apoptosis in tumor cells in a similar manner to that of curcumin. Strikingly, miR-142-5p overexpression suppressed crizotinib-induced autophagy in A549 and H460 cells. Mechanistically, miR-142-5p inhibited autophagy in lung cancer cells by targeting Ulk1. Overexpression of Ulk1 abrogated the miR-142-5p-induced elevation of crizotinib cytotoxicity in A549 and H460 cells. Collectively, our findings demonstrate that curcumin sensitizes NSCLC cells to crizotinib by inactivating autophagy through the regulation of miR-142-5p and its target Ulk1.

miR-365a-5p suppresses gefitinib resistance in non-small-cell lung cancer through targeting PELI3.

Aim: Demonstrate the function of dysregulated miR-365a-5p-PELI3 signaling axis in the generation of gefitinib resistance during treatment for non-small-cell lung cancer (NSCLC). Patients & methods: All the NSCLC patients who participated in this research were recruited from the Second Hospital of Hebei Medical University. PC9 cells and PC9GR cells were cultured for in vitro experiments. Results: Patients who were primary resistant to EGFR-tyrosine kinase inhibitor had lower miR-365a-5p levels. MiR-365a-5p directly targeted PELI3 mRNA. MiR-365a-5p overexpression enhanced the function of gefitinib in inhibiting cell viability. Tumor growth was suppressed through miR-365a-5p in nude mice. Conclusion: Dysregulated miR-365a-5p-PELI3 signaling axis triggered the generation of gefitinib resistance in NSCLC.

Understanding the Rising Phase of the PM2.5 Concentration Evolution in Large China Cities.

Long-term air quality observations are seldom analyzed from a dynamic view. This study analyzed fine particulate matter (PM2.5) pollution processes using long-term PM2.5 observations in three Chinese cities. Pollution processes were defined as linearly growing PM2.5 concentrations following the criteria of coefficient of determination R2 > 0.8 and duration time T ≥ 18 hrs. The linear slopes quantitatively measured pollution levels by PM2.5 concentrations rising rates (PMRR, µg/(m3·hr)). The 741, 210 and 193 pollution processes were filtered out, respectively, in Beijing (BJ), Shanghai (SH), and Guangzhou (GZ). Then the relationships between PMRR and wind speed, wind direction, 24-hr backward points, gaseous pollutants (CO, NO2 and SO2) concentrations, and regional PM2.5 levels were studied. Inverse relationships existed between PMRR and wind speed. The wind directions and 24-hr backward points converged in specific directions indicating long-range transport. Gaseous pollutants concentrations increased at variable rates in the three cities with growing PMRR values. PM2.5 levels at the upwind regions of BJ and SH increased at high PMRRs. Regional transport dominated the PM2.5 pollution processes of SH. In BJ, both local contributions and regional transport increased during high-PMRR pollution processes. In GZ, PM2.5 pollution processes were mainly caused by local emissions.

Daily estimation of ground-level PM2.5 concentrations at 4km resolution over Beijing-Tianjin-Hebei by fusing MODIS AOD and ground observations.

The satellite-borne Moderate Resolution Imaging Spectroradiometer (MODIS) aerosol optical depth (AOD) is widely used to estimate ground-level fine ambient particulate matter (PM2.5) concentrations to evaluate their health effects. The associated estimation accuracy is often reduced by AOD missing values and by insufficiently accounting for the spatio-temporal PM2.5 variations. In this study, we aim to estimate ground-level PM2.5 concentrations at a fine resolution with improved accuracy by fusing fine-scale satellite and ground observations in the populated and polluted Beijing-Tianjin-Hebei (BTH) area of China in 2014. We employed a Bayesian-based statistical downscaler to model the spatio-temporal linear AOD-PM2.5 relationships. We used a 3km MODIS AOD product, which was resampled to a 4km resolution in a Lambert conic conformal projection, to assist comparison and fusion with predictions by atmospheric chemistry models. A two-step method was used to fill the missing AOD values to obtain a full AOD dataset with complete spatial coverage. The downscaler has a good performance in the fitting procedure (R2=0.75) and in the cross validation procedure (R2=0.58 by random method and R2=0.47 by city-specific method). The number of missing AOD values was serious and related to elevated PM2.5 concentrations. The gap-filled AOD values corresponded well with our understanding of PM2.5 pollution conditions in BTH. The prediction accuracy of PM2.5 concentrations were improved in terms of their annual and seasonal mean. As a result of its fine spatio-temporal resolution and complete spatial coverage, the daily PM2.5 estimation dataset could provide extensive and insightful benefits to related studies in the BTH area. This may include understanding the formation processes of regional PM2.5 pollution episodes, evaluating daily human exposure, and establishing pollution controlling measures.

MicroRNA-148b is a potential prognostic biomarker and predictor of response to radiotherapy in non-small-cell lung cancer.

miR-148b has been found to be aberrantly expressed in various tumor types. It has recently been reported to be involved in regulating radioresistance in non-small cell lung cancer (NSCLC) cells. However, its expression level and association with radiosensitivity in human patient samples have not been investigated. Real-time PCR was used to evaluate the expression levels of miR-148b. Χ (2) test was performed to analyze the association between miR-148b expression levels and clinicopathological factors or radiosensitivity. Kaplan-Meier survival curve was constructed to estimate the overall survival (OS), and the differences in survival were compared using the log-rank test. Cox regression analysis was conducted to determine the prognostic value of miR-148b. The relative level of miR-148b was significantly decreased in NSCLC tissues compared with matched non-cancerous tissues (P < 0.0001), and it was higher in tissues of patients who are good responders compared to those who are poor responders to radiotherapy (P < 0.0001). Lower expression of miR-148b was positively associated with high tumor stage (P = 0.0407) and radioresistance (P = 0.0002), and it predicted poor survival in patients with (P = 0.0129) or without (P = 0.0094) radiotherapy treatment. miR-148b was an independent prognostic factor for NSCLC as demonstrated by Cox proportional hazards risk analysis. miR-148b may serve as a prognostic biomarker of poor survival and a novel predictor of response to radiotherapy treatment in NSCLC.

microRNA-214 Governs Lung Cancer Growth and Metastasis by Targeting Carboxypeptidase-D.

Lung cancer is one of the most malignant cancers with a high metastatic potential. The purpose of this study was to study the role and the underlying mechanism of miR-214 in lung cancer progression. The expression of miR-214 in normal lung and lung cancer tissue was analyzed by quantitative real-time PCR analysis. Furthermore, H1299 cells were infected with miR-214 lentivirus, and the effect of infection on cell viability and migration was analyzed. Carboxypeptidase-D (CPD), as a potential target of miR-214, was characterized in either normal lung or lung cancer tissues. The interaction of CPD expression with the tumor suppressing effect of miR-214 was characterized. We demonstrated that low miR-214 expression is a hallmark of lung cancer, especially high-grade and metastatic cancer. In vitro studies in H1299 cells confirmed that low miR-214 expression is associated with enhanced proliferation and migratory abilities. Similarly, CPD overexpression coincides with high-grade lung cancer and the CPD overexpression could reverse the inhibitory effects of miR-214. miR-214 is a tumor suppressor in lung cancer. miR-214 inhibits lung cancer progression by targeting CPD. The miR-214-CPD axis may be a therapeutic axis for lung cancer patients.

Endothelial PAS domain-containing protein 1 confers TKI-resistance by mediating EGFR and MET pathways in non-small cell lung cancer cells.

Mutations in epidermal growth factor receptor (EGFR) rendering it constitutively active is one of the major causes for metastatic non-small-cell lung cancer (NSCLC), and EGFR-targeted therapies utilizing tyrosine kinase inhibitors (TKIs) are often used clinically as the first-line treatment. But approximately half of NSCLC patients develop resistance to these therapies, where the MET proto-oncogene is amplified by EGFR through the hypoxia-inducible factor (HIF)-1α. Here we report that endothelial PAS domain-containing protein 1 (EPAS1), with 48% sequence identity to HIF-1α, specifically binds to TKI-resistant T790M EGFR, but not to wild-type EGFR, in NSCLC cell lines. Expression of EPAS1 enhances amplification of MET when simultaneously expressed with T790M EGFR but not with wild-type EGFR, and this enhancement is independent of ligand binding domain of EGFR. MET amplification requires EPAS1, since EPAS1 knock-down reduced MET levels. When NSCLC cells expressing T790M EGFR were treated with TKIs, reduced EPAS1 levels significantly enhanced the drug effect, whereas over-expression of EPAS1 increased the drug resistant effect. This EPAS1-dependent TKI-resistance was abolished by knocking-down MET, suggesting that EPAS1 does not cause TKI-resistance itself but functions to bridge EGFR and MET interactions. Our findings suggest that EPAS1 is a key factor in the EGFR-MET crosstalk in conferring TKI-resistance in NSCLC cases, and could be used as a potential therapeutic target in TKI-resistant NSCLC patients.