TMEM9 promotes lung adenocarcinoma progression via activating the MEK/ERK/STAT3 pathway to induce VEGF expression.

Abnormal Transmembrane protein 9 (TMEM9) expression has been identified in various human tumors. However, the prognostic potential and mechanistic role of TMEM9 in lung adenocarcinoma (LUAD) remain unclear. Here, we first found a significant upregulation of TMEM9 in LUAD tissues, and TMEM9 expression was positively correlated with microvessel density (MVD), T stage, and clinical stage. Survival analysis demonstrated TMEM9 was an independent indicator of poor prognosis in LUAD patients. In addition, downregulation of TMEM9 suppressed tumor growth and metastasis in vitro and in vivo models, and reduced HUVEC proliferation, migration, and tube formation in a cancer cell/HUVEC coculture model. Furthermore, TMEM9 upregulated VEGF expression, and VEGF-neutralizing antibodies reversed HUVEC angiogenesis and cancer cell migration ability caused by overexpression of TMEM9. In contrast, recombinant VEGF (rVEGF) abolished the inhibitory effect of TMEM9-knockdown LUAD cells on HUVEC angiogenesis and tumor cell migration. Moreover, we showed that TMEM9 upregulated VEGF expression by activating the mitogen-activated protein kinase/extracellular signal-regulated kinase/STAT3 (MEK/ERK/STAT3) pathway. Together, our study provides mechanistic insights into the role of TMEM9 in LUAD and highlights the potential of targeting the TMEM9/MEK/ERK/STAT3/VEGF pathway as a novel therapy for preventing LUAD progression.

Genomic Features of Organ-Specific Metastases in Lung Adenocarcinoma.

Background: The genomic features of cancer cells may confer the metastatic ability of lung adenocarcinoma (LUAD) to metastasize to specific organs. We aimed to identify the differences in genomic alterations between patients with primary LUAD with and without metastases and to elucidate the metastatic biology that may help developing biomarker-directed therapies for advanced or metastatic disease. Methods: A retrospective cohort of 497 patients with LUAD including 388 primary tumors (PR), 53 bone metastases (MT-bone), 30 liver metastases (MT-liver), and 26 brain metastases (MT-brain) was tested for genomic alterations by a next-generation sequencing assay. Results: The EGFR, TP53, TERT, LRP1B, CDKN2A, ERBB2, ALK, and KMT2C genes had a high frequency of mutations, and the mutations were shared by PR and metastases groups. TP53 and EGFR were the most common mutated genes. In comparison with PR, KRAS, STK11, ATM, NPM1, and ROS1 were significantly mutated in MT-brain, and TP53, MYC, RSPO2, CDKN2a, and CDKN2B were significantly mutated in MT-liver. The frequencies of TP53, CDKN2A, MTAP, PRKCI, and APC mutations were higher in MT-bone than that in PR. The ERBB, phosphoinositide-3-kinase/protein kinase B (PI3K-AKT), cell cycle, Fibroblast growth factor (FGF), and homologous recombination deficiency signaling pathways were affected in both PR and metastases, and there is higher frequency of mutations in metastases. Moreover, the co-mutations in patients with PR and metastasis were respectively analyzed. In addition, the programmed death ligand 1 (PD-L1) level was obviously related to tumor stage and tumor metastases, and the tumor mutational burden was correlated to clinicopathological features including age, gender, pathological stages, and tumor metastases. FGFR1, KAT6A, MYC, RAD21, TP53, and DAXX were also dramatically correlated to the tumor mutational burden. Conclusion: Metastases are the most devastating stage of tumors and the main cause of cancer-related deaths. Our results provided a clinically relevant view of the tumor-intrinsic mutational landscape of patients with metastatic LUAD.

Positron emission tomography/computed tomography and endobronchial ultrasound-guided transbronchial needle aspiration to evaluate the status of N2 in preoperative non-small cell lung cancer: a diagnostic test.

Background: As a minimally invasive method, endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) was more accurate than non-invasive methods such as positron emission tomography (PET) and computed tomography (CT) to evaluate the lymph nodes in preoperative non-small cell lung cancer (NSCLC). PET/CT has more anatomical advantages than PET scanning and is more accurate in lung cancer staging. However, no relevant studies have comparatively evaluated PET/CT and EBUS-TBNA for NSCLC patients. Methods: A total of 112 patients were included in this retrospective analysis. The golden diagnosis of N2 status was postoperative pathological results. In EBUS-TBNA puncture specimens, if clear malignant tumor cells could be seen, the results were taken as positive. In PET/CT image analysis, the CT values, short diameter, and maximum standardized uptake value (SUVmax) of each lymph node were recorded to evaluate N2 status. The results of PET/CT and EBUS-TBNA were compared with the final pathological results, and respective sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and accuracy were calculated. - Then, the patients were divided into adenocarcinoma group and squamous cell carcinoma group -and the results were calculated and compared with the above method. Results: The results showed that EBUS-TBNA had a higher diagnostic value for mediastinal lymph nodes than PET/CT, and the difference was statistically significant (P<0.001). In NSCLC patients, the results showed that the sensitivity (P=0.013), specificity (P<0.001), PPV (P<0.001), NPV (P<0.001), and accuracy (P<0.001) of EBUS-TBNA were higher than that of PET/CT (AUC =0.954 and 0.636, respectively). In adenocarcinoma cases, specificity (P<0.001), PPV (P<0.001), NPV (P<0.001), and accuracy (P<0.001) of EBUS-TBNA were higher than that of PET/CT (AUC =0.957 and 0.596, respectively).In cases with squamous cell carcinoma, specificity (P=0.003), PPV (P<0.001), and accuracy (P<0.001) of EBUS-TBNA were higher than PET/CT (AUC =0.952 and 0.657, respectively). Conclusions: For preoperative diagnosis of mediastinal lymph node metastases in NSCLC, EBUS-TBNA is more accurate than PET/CT. For those patients with suspected mediastinal lymph node metastasis, EBUS-TBNA should be preferred method to evaluate the status of mediastinal lymph nodes.

Circular RNA hsa_circ_0077837 is upregulated in non-small cell lung cancer to downregulate phosphatase and tensin homolog through methylation.

Circular RNA (circRNA) hsa_circ_0077837 inhibits colorectal cancer. Our research studied the participation of hsa_circ_0077837 in non-small cell lung cancer (NSCLC). Hsa_circ_0077837 and phosphatase and tensin homolog (PTEN) expression in cancer and paired non-cancer tissues from a total of 64 NSCLC patients were studied with RT-qPCR. To evaluate the prognostic value of hsa_circ_0077837 for NSCLC, these 64 patients were monitored for 5 years. Expression of PTEN in NSCLC cells with hsa_circ_0077837 overexpression was determined by RT-qPCR and Western blot. The methylation of PTEN gene in cells transfected with hsa_circ_0077837 expression vector was analyzed by methylation specific PCR (MSP). The roles of hsa_circ_0077837 and PTEN in NSCLC cell proliferation were evaluated using cell apoptosis assay. Our data showed that hsa_circ_0077837 was upregulated in NSCLC and predicted poor survival. Besides, hsa_circ_0077837 expression level was higher in 36 advanced cases (stage III and IV) than in 28 early-stage cases (stage I and II). Hsa_circ_0077837 was inversely correlated with PTEN across cancer tissues. In NSCLC cells, hsa_circ_0077837 overexpression decreased PTEN expression, increased PTEN gene methylation, and reduced HCC827 cell apoptosis via PTEN. Overall, hsa_circ_0077837 is upregulated in NSCLC and downregulates PTEN by increasing its gene methylation to suppress cell apoptosis.List of abbreviations:Non-small cell lung cancer (NSCLC); circRNAs (circular RNAs); methylation-specific PCR (MSP).

The role of ERBB4 mutations in the prognosis of advanced non-small cell lung cancer treated with immune checkpoint inhibitors.

BACKGROUND: Immune checkpoint inhibitors (ICIs) have witnessed the achievements of convincing clinical benefits that feature the significantly prolonged overall survival (OS) of patients suffering from advanced non-small cell lung cancer (NSCLC), according to reports recently. Sensitivity to immunotherapy is related to several biomarkers, such as PD-L1 expression, TMB level, MSI-H and MMR. However, a further investigation into the novel biomarkers of the prognosis on ICIs treatment is required. In addition, there is an urgent demand for the establishment of a systematic hazard model to assess the efficacy of ICIs therapy for advanced NSCLC patients. METHODS: In this study, the gene mutation and clinical data of NSCLC patients was obtained from the TCGA database, followed by the analysis of the detailed clinical information and mutational data relating to two advanced NSCLC cohorts receiving the ICIs treatment from the cBioPortal of Cancer Genomics. The Kaplan-Meier plot method was used to perform survival analyses, while selected variables were adopted to develop a systematic nomogram. The prognostic significance of ERBB4 in pan-cancer was analyzed by another cohort from the cBioPortal of Cancer Genomics. RESULTS: The mutation frequencies of TP53 and ERBB4 were 54% and 8% in NSCLC, respectively. The mutual exclusive analysis in cBioPortal has indicated that ERBB4 does show co-occurencing mutations with TP53. Patients with ERBB4 mutations were confirmed to have better prognosis for ICIs treatment, compared to those seeing ERBB4 wild type (PFS: exact p = 0.017; OS: exact p < 0.01) and only TP53 mutations (OS: p = 0.021). The mutation status of ERBB4 and TP53 was tightly linked to DCB of ICIs treatment, PD-L1 expression, TMB value, and TIICs. Finally, a novel nomogram was built to evaluate the efficacy of ICIs therapy. CONCLUSION: ERBB4 mutations could serve as a predictive biomarker for the prognosis of ICIs treatment. The systematic nomogram was proven to have the great potential for evaluating the efficacy of ICIs therapy for advanced NSCLC patients.

[A Review of Epigenetic Modifications Regulate MicroRNA Expression in Lung Cancer].

Lung cancer is one of the primary causes of cancer-induced death among the world. Although the network of molecular implicated in lung cancer is gradually revealed, the exact molecular mechanism of its occurrence and development has not been fully elucidated. As a class of small and endogenous single-stranded non-coding RNAs, microRNAs (miRNAs) are found in a wide range of organisms from plants, viruses to humans. miRNAs involve various functions in normal lung tissue development. They take part in a large amount of biological processes including cell growth, metabolism, proliferation and differentiation. However, aberrant expression of miRNAs could induce the occurrence, development, invasion and metastasis of lung tumor, so it is deemed the novel biomarkers. Similar to that of protein-coding genes, expression and function of miRNA are regulated by various factors and the epigenetic network which includes DNA methylation and histone modification. Moreover, key enzymes driving epigenetic modifications are regulated by miRNAs. Therefore, better understanding of inextricable linkage between miRNAs and epigenome will provides a basis for the feasibility of miRNA-orientated diagnostic, therapeutic and prognostic strategies related to lung cancer in future.

Investigation of the effect of P14 promoter aberrant methylation on the biological function of human lung cancer cells.

BACKGROUND: This study was conducted to investigate the effect of P14 promoter aberrant methylation on the biological function of human lung adenocarcinoma cells. METHODS: We used nested methylation-specific PCR (NMSP) to detect the methylation status of the p14ARF promoter region in SPCA1 and BEAS2B cell lines. The experimental groups were treated with 5-aza-2'-deoxycytidine (5-Aza). Quantitative real-time PCR, Western blot, flow cytometry, and Cell Counting Kit 8 were used to detect the expression of p14ARF messenger RNA and protein in each group, apoptosis, and cell proliferation inhibition, respectively. RESULTS: NMSP detected that the p14 promoter region of SPCA1 cells has abnormal methylation status. After treatment with 5-Aza, the expression of p14ARF messenger RNA and protein in SPCA1 cells (P < 0.05) and the inhibition rate of cell proliferation (P < 0.05) were significantly increased, while the apoptosis rate was markedly increased (P < 0.05). However, no differences were observed in BEAS2B cells (P > 0.05). CONCLUSION: Abnormal methylation of the p14ARF promoter region plays an important role in the development of lung cancer cells. Our results suggest the use of P14 promoter aberrant methylation as a therapeutic target for drug research or to improve the sensitivity of other drugs.

Left upper lobectomy and pulmonary angioplasty by uniportal video-assisted thoracic surgery.

Diego Gonzalez-Rivas and his colleagues performed the first case for pulmonary major resection by the uniportal approach in 2010. In the following years, more and more expert surgeons developed the uniportal approach and even applied it to very complex cases. Many centers have showed that uniportal video-assisted thoracic surgery (VATS) lobectomy is a safe and feasible technique. The patient presented here is a complex case: The lingular segmental artery was invaded seriously by the tumor and it was difficult to divide it freely, and the pulmonary artery plasty was needed. Although this condition made the procedure some technically challenging, the case was performed successfully without any operative complications.

The decreased expression of miR-625 predicts poor prognosis of esophageal squamous cell carcinoma.

BACKGROUND: Previous study has detected the expression of miR-625 in esophageal squamous cell carcinoma (ESCC) and found that miR-625 was related to tumor depth, stage, and metastasis of ESCC. However, the prognostic value of miR-625 in ESCC has not yet been reported. METHODS: Real-time quantitative PCR was employed to measure the expression level of miR-625 in clinical ESCC tissues. Survival curves were made using the Kaplan-Meier method, and the log rank test was used to analyze the differences between clinicopathological characteristics and survival in ESCC patients. RESULTS: The expression level of miR-625 in ESCC tissues was significantly lower than that in adjacent non-tumor tissues (1.00 ± 0.38 vs. 3.25 ± 1.83, P < 0.0001). Low miR-625 expression was observed to be closely correlated with lymph node metastasis (P = 0.01), distant metastasis (P = 0.007), tumor differentiation (P = 0.04), and advanced TNM stage (P = 0.005). The 5-year overall survival rate in the low expression group was 38.1%, compared with 68.8% in the high expression group (log-rank test, P = 0.001). Multivariate Cox regression analysis showed that miR-625 expression level (HR = 3.72, 95% CI: 1.36-8.78, P = 0.005) was an independent factor in predicting the overall survival of ESCC patients. CONCLUSION: Our findings provide the convincing evidence for the first time that the down-regulation of miR-625 may serve as a novel molecular marker to predict the aggressive tumor progression and unfavorable prognosis of ESCC patients.

Investigation on immune function and chest drainage in patients with thoracic malignancies using the streptococcal agent Sapylin.

PURPOSE: To investigate the influence of Sapylin used during the operation on immune function and chest drainage in patients with thoracic malignancies in early postoperative period. PATIENTS AND METHODS: Seventy-one patients with thoracic malignancies including lung cancer and esophageal carcinoma who underwent tumor resection were divided into two groups, 36 cases of observation group and 35 cases of control group. In observation group, Sapylin were locally laid in operation field during the operation. The control group was not given Sapylin. Drainage volume of chest cavity after operation, and the levels of CD4, CD8, CD4/CD8 and natural killer (NK) cell were recorded. RESULTS: The level of CD4+, CD4+/CD8+ and NK cell in observation group were higher than that in control group (P<0.05) wholly in early postoperative period. Drainage volume of chest cavity in observation group was less than that in control group postoperatively, 1160.2±437.2 ml vs. 1363.3±472.9 ml, but the change in two groups had no significance statistically (P=0.064). CONCLUSIONS: The local application of Sapylin during operation can improve immunity of patients with thoracic malignancies significantly and decrease the drainage volume of chest cavity probably in early postoperative period. So it is worthy of popularizing in clinical practice.

[Promoter methylation status and protein expression of p14ARF gene in squamous cell carcinoma and adenocarcinoma of the lung].

BACKGROUND: Recently, the p14 ARF gene has emerged as a new putative tumor suppressor gene, and the alteration of p14 ARF gene is closely related to development of multiple human tumors. The aberrant promoter methylation as a mechanism of inactivation of p14 ARF gene might participate in tumorigenesis. The aim of this study is to investigate promoter methylation status and protein expression of p14 ARF gene in pulmonary squamous cell carcinoma and adenocarcinoma, and to value the role of p14 ARF promoter methylation in carcinogenesis of non-small cell lung cancer. METHODS: Promoter methylation status and protein expression of p14 ARF gene were analyzed in 40 cases of pulmonary squamous cell carcinoma and adenocarcinoma by methylation specific polymerase chain reaction (MSP), restriction enzyme-related polymerase chain reaction (RE-PCR) and immunohistochemistry (IHC). RESULTS: The positive rate of p14 ARF promoter methylation in tumor tissues and normal tissues adjacent to cancer was 17.5% (7/40) and 2.5% (1/40) respectively (P= 0.025 ). The results of RE-PCR were consistent with the above. The positive rate of p14 ARF protein in tumor tissues was significantly lower than that in normal tissues adjacent to cancer (P=0.003). Promoter methylation and protein expression of p14 ARF gene showed a significantly negative correlation (r=-0.56, P= 0.001 ), and both of them did not correlate statistically with the clinicopathologic characteristics of patients such as histological classification, TNM stages, differentiation grade and lymph node involvement. CONCLUSIONS: Promoter methylation is a crucial mechanism of inactivation of p14 ARF gene. Promoter methylation of p14 ARF gene might be involved in carcinogenesis of non-small cell lung cancer, and it is an early event in development process of non-small cell lung cancer.