Transcription Factor MITF Inhibits the Transcription of CPT1B to Regulate Fatty Acid ß-Oxidation and Thus Affects Stemness in Lung Adenocarcinoma Cells.

INTRODUCTION: Cancer stem cells (CSCs) play critical roles in lung adenocarcinoma (LUAD) progression, and fatty acid oxidation is key for CSC growth and survival. Therefore, investigating the molecular mechanisms regulating fatty acid ß-oxidation in LUAD is important for its treatment. METHODS: Bioinformatics analysis assessed CPT1B and MITF expression and their correlation in LUAD tissues, as well as the pathways enriched by CPT1B. qRT-PCR assessed expression of CPT1B and MITF, while CCK-8 and sphere-forming assays were used to measure cell viability and stemness, respectively. Dual staining detected lipid accumulation, while kits were used to measure fatty acid ß-oxidation and glycerol content. qRT-PCR was used to assay expression of lipid oxidation genes. Western blot was used to examine expression of stem cell-related markers. Dual-luciferase assay and ChIP assay were used to verify the binding relationship between MITF and CPT1B. RESULTS: CPT1B was found to be highly expressed in LUAD and enriched in linoleic acid metabolism pathway and α-linolenic acid metabolism pathway. Functional experiments showed that CPT1B could promote stemness in LUAD cells by regulating fatty acid ß-oxidation. Additionally, CPT1B was found to be regulated by the upstream transcription factor MITF, which was lowly expressed in LUAD and could downregulate CPT1B expression. Rescue experiments revealed that CPT1B/MITF axis could affect stemness in LUAD cells by regulating fatty acid ß-oxidation. CONCLUSION: Transcription factor MITF inhibited transcription of CPT1B to regulate fatty acid ß-oxidation, thereby suppressing stemness in LUAD cells. MITF and CPT1B may become new targets for LUAD.

TRPM7 overexpression enhances the cancer stem cell-like and metastatic phenotypes of lung cancer through modulation of the Hsp90α/uPA/MMP2 signaling pathway.

BACKGROUND: Waixenicin A, a bioactive extract of soft coral Sarcothelia edmondsoni, has been shown to be anti-neoplastic. However, its mechanisms of action remain unclear. Cancer stem cells (CSCs) and associated stemness factors are implicated in lung cancer. Here, we investigated the role of Waixenicin A on CSCs-like and metastatic lung cancer cells. METHODS: We demonstrated and compared TRPM7 expression in the non-tumor lung tissues or bronchial epithelial 16-HBE cell line. TRPM7 was aberrantly expressed in the cancer tissues and SPCA-1, NCI-H520, SK-MES-1, A549 and 95D cell lines. RESULTS: Increased TRPM7 expression was associated with enhanced SOX2, KLF4, and CD133, Hsp90α, uPA, and MMP2 expression in lung cancer cells. TRPM7-silencing inhibited epithelial-to-mesenchymal transition (EMT), suppressed stemness markers and phenotypes, concomitantly suppressed Hsp90α/uPA/MMP2 axis. Coincidently, Waixenicin A treatment downregulated TRPM7 and oncogenic markers; Waixenicin A also attenuated the ability of lung cancer cells to form tumorspheres, in vitro. In validation, our clinicopathological analyses showed that a higher TRPM7 expression was positively correlated with the larger tumor size (p = 0.007), positive lymph node metastasis (p = 0.005) and disease grade (p = 0.003). CONCLUSIONS: Through its ability to inhibit Hsp90α/uPA/MMP2 signaling and suppress TRPM7 expression, we showed that Waixenicin A is a potential anticancer therapeutic agent for treating malignant lung cancer.

A study on cellular immune function of patients treated with radical resection of pulmonary carcinoma with two different methods of anesthesia and analgesia.

PURPOSE: To compare the influence on the immune system of two different methods of anesthesia and analgesia in patients treated with radical resection of pulmonary carcinoma. METHODS: Thirty-four patients treated with radical resection of pulmonary carcinoma were randomly divided into two groups (group A and group B, 17 cases in each group). Patients in group A were administered total intravenous anesthesia (TIVA) without inhaled hypnotics and intravenous analgesia while patients in group B were administered TIVA combined with epidural anesthesia and epidural analgesia. We compared changes of the T cells subsets (CD3+, CD4+, CD8+ and CD4+/CD8+ ratio) and the function of natural killer (NK) cells in patients at 4 time points: before anesthesia, immediately after surgery, 24 hrs after surgery and 72 hrs after surgery. Clinical data were also collected. RESULTS: CD8+ in group A and B was significantly increased (p<0.01) while the other indexes CD3+, CD4+, CD4+/CD8+ ratio and NK cells) were significantly decreased (p<0.05). There was a significant difference in various indexes (except NK cell) before anesthesia and 72 hrs after surgery in group A (p<0.01). Various indexes of patients in group B at 72 hrs after surgery were restored to the values before anesthesia (p>0.05). We observed a significant difference in CD3+, CD8+ and CD4+/CD8+ indexes in groups A and B patients at 72 hrs after surgery (p<0.05). CONCLUSIONS: TIVA combined with epidural anesthesia and epidural analgesia demonstrated less interference with the immune system and determine fast recovery in patients with radical resection of pulmonary carcinoma.

In silico discovery of potential PPI inhibitors for anti-lung cancer activity by targeting the CCND1-CDK4 complex via the P21 inhibition mechanism.

Non-Small Cell Lung Cancer (NSCLC) is a prevalent and deadly form of lung cancer worldwide with a low 5-year survival rate. Current treatments have limitations, particularly for advanced-stage patients. P21, a protein that inhibits the CCND1-CDK4 complex, plays a crucial role in cell proliferation. Computer-Aided Drug Design (CADD) based on pharmacophores can screen and design PPI inhibitors targeting the CCND1-CDK4 complex. By analyzing known inhibitors, key pharmacophores are identified, and computational methods are used to screen potential PPI inhibitors. Molecular docking, pharmacophore matching, and structure-activity relationship studies optimize the inhibitors. This approach accelerates the discovery of CCND1-CDK4 PPI inhibitors for NSCLC treatment. Molecular dynamics simulations of CCND1-CDK4-P21 and CCND1-CDK4 complexes showed stable behavior, comprehensive sampling, and P21's impact on complex stability and hydrogen bond formation. A pharmacophore model facilitated virtual screening, identifying compounds with favorable binding affinities. Further simulations confirmed the stability and interactions of selected compounds, including 513457. This study demonstrates the potential of CADD in optimizing PPI inhibitors targeting the CCND1-CDK4 complex for NSCLC treatment. Extended simulations and experimental validations are necessary to assess their efficacy and safety.

HES6 Mediates Oxidative Phosphorylation Pathway to Promote Immune Infiltration of CD8 + T Cells in Lung Adenocarcinoma.

Tumor immunotherapy has recently gained popularity as a cancer treatment strategy. The molecular mechanism controlling immune infiltration in lung adenocarcinoma (LUAD) cells, however, is not well characterized. Investigating the immune infiltration modulation mechanism in LUAD is crucial. LUAD patient samples were collected, and HES6 expression and immune infiltration level of CD8 + T cells in patient tissues were analyzed. Bioinformatics was utilized to identify binding relationship between E2F1 and HES6, and enrichment pathway of HES6. The binding of E2F1 to HES6 was verified using dual-luciferase and ChIP experiments. HES6 and E2F1 expression in LUAD cells was detected. LUAD cells were co-cultured with CD8 + T cells, and the CD8 + T cell killing level, IFN-γ secretion, and CD8 + T-cell chemotaxis level were measured. Expression of key genes involved in oxidative phosphorylation was detected, and the oxygen consumption rate of LUAD cells was assessed. A mouse model was constructed to assay Ki67 expression and apoptosis in tumor tissue. High expression of HES6 promoted CD8 + T-cell infiltration and enhanced T-cell killing ability through oxidative phosphorylation. Further bioinformatics analysis, molecular experiments, and cell experiments verified that E2F1 negatively regulated HES6 by oxidative phosphorylation, which suppressed CD8 + T-cell immune infiltration. In addition, in vivo assays illustrated that silencing HES6 repressed tumor cell immune evasion. E2F1 inhibited HES6 transcription, thereby mediating oxidative phosphorylation to suppress immune infiltration of CD8 + T cells in LUAD. The biological functions and signaling pathways of these genes were analyzed, which may help to understand the possible mechanisms regulating immune infiltration in LUAD.

Benefits and Relevant Risk Factor Assessment of Neoadjuvant Chemotherapy in Combination with Surgery in Limited-Stage Small-Cell Lung Cancer.

To illustrate the benefits of surgery in conjunction with neoadjuvant chemotherapy in patients with limited-stage small cell lung cancer (LS-SCLC), and to evaluate risk factors affecting patient's survival. Forty-six LS-SCLC patients who received surgery in our center from September 2012 to December 2018 were retrospectively analyzed. Twenty-five patients with LS-SCLC diagnosed after surgery who received postoperative adjuvant chemotherapy were classified into control group, and 21 patients with LS-SCLC who received preoperative neoadjuvant chemotherapy were classified into observation group. The observation group were divided into subgroup 1 (negative lymph nodes) and subgroup 2 (positive lymph nodes). Progression-free survival (PFS) and overall survival (OS) of patients were analyzed. Univariate and multivariate Cox regression were utilized to analyze independent risk factors affecting patient's survival. PFS and OS of patients in the control group and observation group had similar outcomes (P > 0.05). Subgroup 1 and subgroup 2 had similar PFS and OS (P > 0.05). PT2, pN2, BM, and two or more positive lymph nodes were significantly associated with poor PFS and OS (P < 0.05). Furthermore, the pT, number of lymph node positive stations and BM were independent risk factors affecting patient's survival (P < 0.05). Surgery combined with neoadjuvant chemotherapy can achieve long-term survival benefit for some patients with LS-SCLC. It is necessary to find a better plan that enables to select patients suitable for surgery after neoadjuvant chemotherapy.

Corrigendum: LncRNA SOX2-OT/miR-30d-5p/PDK1 Regulates PD-L1 Checkpoint Through the mTOR Signaling Pathway to Promote Non-Small Cell Lung Cancer Progression and Immune Escape.

[This corrects the article DOI: 10.3389/fgene.2021.674856.].

ADAMTS9-AS2 regulates PPP1R12B by adsorbing miR-196b-5p and affects cell cycle-related signaling pathways inhibiting the malignant process of esophageal cancer.

This study explored the mechanism that ADAMTS9-AS2/miR-196b-5p/PPP1R12B/cell cycle pathway axis in inhibiting the malignant progression of esophageal cancer (EC), providing a new idea for targeted molecular therapy of EC. The expression data of EC tissue were acquired from TCGA database. The target lncRNA, downstream miRNA and its target gene were determined by bioinformatics analysis. ADAMTS9-AS2, miR-196b-5p and PPP1R12B levels in EC tissue and cells were assayed through qRT-PCR. Western blot was applied to assess protein level of PPP1R12B in cells and tissues, as well as protein expression of CDK1, cyclin A2, cyclin B1 and Plk1 in EC cells. Cell proliferation was assayed via CCK-8 assay. Cell cycle distribution was analyzed by flow cytometry. Cell migratory and invasive abilities were measured through scratch healing and transwell assays. Pearson correlation analysis was utilized to analyze relationship among ADAMTS9-AS2, miR-196b-5p and PPP1R12B. RIP was introduced to assess binding among the three. Dual-luciferase assay was utilized to verify targeted binding sites. The tumor formation in nude mice assay was utilized to detect tumorigenesis of EC cells in vivo. ADAMTS9-AS2 was significantly lowly expressed while miR-196b-5p was increased in EC tissue and cells. ADAMTS9-AS2 bound to miR-196b-5p and constrained its expression. Overexpressed ADAMTS9-AS2 inhibited EC cell malignant progression via downregulating miR-196b-5p, while overexpressed miR-196b-5p reversed this inhibitory effect. ADAMTS9-AS2 modulated PPP1R12B level by competitively inhibiting miR-196b-5p. PPP1R12B played a modulatory role in EC by inhibiting cell cycle pathway. Overexpressed ADAMTS9-AS2 regulated the tumor-forming ability of EC cells in vivo through miR-196b-5p/PPP1R12B/cell cycle signaling pathway axis. ADAMTS9-AS2 downregulated PPP1R12B by adsorbing miR-196b-5p, so as to regulate the cell cycle signaling pathway to inhibit EC malignant progression.

miR-1-3p/CELSR3 Participates in Regulating Malignant Phenotypes of Lung Adenocarcinoma Cells.

OBJECTIVE: This study presents a discussion regarding the mechanism affecting the malignant progression of LUAD and the potential therapeutic targets, so as to provide more effective therapeutic strategies for LUAD patients. METHODS: Expression data from TCGA-LUAD were extracted to identify target miRNA, with its downstream target mRNA predicted using bioinformatics analysis. Gene expression in transcript level and protein level were separately examined by qRT-PCR and western blot. Cell malignant phenotypes were assessed via MTT and Transwell assays. Luciferase reporter plasmids carrying target gene sequences were constructed to verify the targeting association between the target miRNA and its downstream mRNA. RESULTS: miR-1-3p showed decreased expression in LUAD. Over-expressing miR-1-3p suppressed cancer cells to proliferate, migrate and invade. CELSR3, directly regulated by miR-1-3p, presented significantly elevated expression in LUAD and could foster LUAD cells to proliferate, migrate and invade. The rescue experiment identified that miR-1-3p-induced inhibition on LUAD cell malignant phenotypes could be reversed by over-expressing CELSR3. CONCLUSION: This study uncovered that miR-1-3p could suppress the malignant phenotypes of LUAD cells by targeting CELSR3, which will help to provide novel therapeutic strategies for LUAD sufferers and new references for the targeted therapy of LUAD.

LncRNA SOX2-OT/miR-30d-5p/PDK1 Regulates PD-L1 Checkpoint Through the mTOR Signaling Pathway to Promote Non-small Cell Lung Cancer Progression and Immune Escape.

Non-small cell lung cancer (NSCLC) is the most common type of lung cancer. Currently, treatment methods generally cause poor prognosis. Therefore, in order to seek new treatment options, we explored the internal mechanism of NSCLC. Firstly, the SOX2-OT/miR-30d-5p/PDK1 axis regulated by lncRNA SOX2-OT was predicted by bioinformatics methods, and the expression of SOX2-OT, miR-30d-5p, and PDK1 mRNA in cells were detected by qRT-PCR while PDK1 protein expression was detected by western blot. The results expressed that in NSCLC, SOX2-OT, and PDK1 were notably overexpressed while miR-30d-5p was markedly under-expressed. The interaction between them was verified by dual-luciferase reporter and RNA binding protein immunoprecipitation assays. Subsequently, through CCK8, scratch healing, cell invasion and flow cytometry assays, we revealed that inhibiting the expression of SOX2-OT could inhibit the proliferation, migration and invasion of NSCLC cells and promote cell apoptosis; while simultaneous overexpression of PDK1 or inhibition of miR-30d-5p expression could reverse the inhibitory effect of SOX2-OT silence-mediated malignant progression of NSCLC cells. Then, the combined application of overexpressed PDK1 and rapamycin verified that PDK1 could regulate the expression of PD-L1 in NSCLC cells through the mTOR signaling pathway. Co-culture of CD8+ T cells verified that silencing SOX2-OT could inhibit the apoptosis of CD8+ T cells through miR-30d-5p/PDK1. Finally, tumor formation assay in animals confirmed that overexpression of SOX2-OT could promote the growth of NSCLC tumor in vivo. In this study, assays in vitro and in vivo were conducted to elucidate the mechanism by which SOX2-OT/miR-30d-5p/PDK1 drives PD-L1 through the mTOR signaling pathway to promote the malignant progression and immune escape of NSCLC.

Andrographolide inhibits non-small cell lung cancer cell proliferation through the activation of the mitochondrial apoptosis pathway and by reprogramming host glucose metabolism.

BACKGROUND: The main aim of this research was to explore the role and mechanism of Andrographolide (Andro) in controlling non-small cell lung cancer (NSCLC) cell proliferation. METHODS: Human NSCLC H1975 cells were treated with Andro (0-20 µM) for 4-72 h. B-cell leukemia/lymphoma 2 (Bcl-2)-antagonist/killer (Bak)-small interfering RNA (siRNA) (Bak-siRNA) and fructose-1,6-bisphosphatase (FBP1)-siRNA were transfected into H1975 cells to inhibit the endogenic Bak and FBP1 expression, respectively, and their expressions were detected by real-time quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and western blotting (WB). Cellular proliferation ability was determined through various assessments, including 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), colony formation, and cell counting kit-8 (CCK-8) assays. Cell apoptosis ability was measured using flow cytometry. Pro-apoptotic-related proteins (cleaved caspase 9, cleaved caspase 8, and cleaved caspase 3) and mitochondrial apoptosis pathway proteins [Bcl2-associated X (Bax), Bak, Bcl-2, and cytochrome C (cyto C)] were assessed by WB. Aerobic glycolysis-associated genes [pyruvate kinase M2 (PKM2), lactate dehydrogenase A (LDHA), and glucose transporter 1 (GLUT1)] and gluconeogenesis genes [phosphoenolpyruvate carboxykinase 1 (PEPCK1), fructose-1,6-bisphosphatase 1 (FBP1), and phosphofructokinase (PFK)] were measured by qRT-PCR. The mitochondrial membrane depolarization sensor, 5, 50, 6, 60-tetrachloro-1, 10, 3, 30 tetraethyl benzimidazolo carbocyanine iodide (JC-1) assay was used for the measurement of mitochondrial membrane potential (ΔΨm). Additionally, glycolytic metabolism, lactate production, and adenosine triphosphate (ATP) synthesis were also analyzed. RESULTS: Andro inhibited human NSCLC cellular proliferation and induced apoptosis in a dose-time or dose-dependent manner via activation of the mitochondrial apoptosis pathway. Andro inhibited glycolysis, promoted the gluconeogenesis pathway, and increased the levels of cleaved caspase 9, cleaved caspase 8, cleaved caspase 3, Bax, Bak, PEPCK1, FBP1, and PFK, and decreased the levels of Bcl-2, PKM2, LDHA, and GLUT1. Moreover, it also decreased the ΔΨm and facilitated the release of cyto C from mitochondria into the cytoplasm. Furthermore, Andro enhanced the mitochondrial translocation of Bak, glucose uptake, lactate release, and intracellular ATP synthesis. Suppression of endogenic Bak and FBP1 expression significantly reduced the effects of Andro in H1975 cells. CONCLUSIONS: Andro represses NSCLC cell proliferation through the activation of the mitochondrial apoptosis pathway and by reprogramming glucose metabolism.

A Recurrence-Specific Gene-Based Prognosis Prediction Model for Lung Adenocarcinoma through Machine Learning Algorithm.

BACKGROUND: After curative surgical resection, about 30-75% lung adenocarcinoma (LUAD) patients suffer from recurrence with dismal survival outcomes. Identification of patients with high risk of recurrence to impose intense therapy is urgently needed. MATERIALS AND METHODS: Gene expression data of LUAD were obtained from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) databases. Differentially expressed genes (DEGs) were calculated by comparing the recurrent and primary tissues. Prognostic genes associated with the recurrence-free survival (RFS) of LUAD patients were identified using univariate analysis. LASSO Cox regression and multivariate Cox analysis were applied to extract key genes and establish the prediction model. RESULTS: We detected 37 DEGs between primary and recurrent LUAD tumors. Using univariate analysis, 31 DEGs were found to be significantly associated with RFS. We established the RFS prediction model including thirteen genes using the LASSO Cox regression. In the training cohort, we classified patients into high- and low-risk groups and found that patients in the high-risk group suffered from worse RFS compared to those in the low-risk group (P < 0.01). Concordant results were confirmed in the internal and external validation cohort. The efficiency of the prediction model was also confirmed under different clinical subgroups. The high-risk group was significantly identified as the risk factor of recurrence in LUAD by the multivariate Cox analysis (HR = 13.37, P = 0.01). Compared to clinicopathological features, our prediction model possessed higher accuracy to identify patients with high risk of recurrence (AUC = 96.3%). Finally, we found that the G2M checkpoint pathway was enriched both in recurrent tumors and primary tumors of high-risk patients. CONCLUSIONS: Our recurrence-specific gene-based prognostic prediction model provides extra information about the risk of recurrence in LUAD, which is conducive for clinicians to conduct individualized therapy in clinic.

Cullin 3 overexpression inhibits lung cancer metastasis and is associated with survival of lung adenocarcinoma.

Cullin 3 (CUL3), a molecular scaffold of Cullin-RING ubiquitin ligase, plays an important role in regulating biological processes through modulating the ubiquitylation and degradation of various protein substrates. Dysfunction of CUL3 is implicated in the development of several human diseases. However, the clinical significance and prognostic value of CUL3 in lung cancer have not been investigated. This study investigated the CUL3-modulating potential of non-small cell lung cancer cell lines, H1299, H358, H2170 and H520, by using immunoblotting, MTT, migration, invasion, colony formation and in vivo tumorigenicity assays. The prognostic significance of CUL3 was measured by public KM plotter database (http://kmplot.com/analysis/index.php?p=service&cancer=breast) and tissue immunohistochemistry analysis. The public online database analysis revealed that elevated mRNA expression of CUL3 was associated with better prognosis for non-small cell lung cancer and lung adenocarcinoma. In vitro experiments showed that ectopic overexpression of CUL3 significantly inhibited lung adenocarcinoma cell proliferation and migration, and the tumor-suppressive effect of CUL3 was dependent on the Nrf2/RhoA axis. In vivo mice model demonstrated that overexpression of CUL3 lead to a significant reduction of lung adenocarcinoma growth and metastasis. Importantly, tissue immunohistochemistry analysis showed that about 47% of non-small cell lung cancer tissues were expressed of CUL3 at high levels. Overexpression of CUL3 predicted favorable overall survival in non-small cell lung cancer patients, especially in lung adenocarcinoma, but not in lung squamous cell carcinoma patients. CUL3 could serve as a prognostic biomarker for lung adenocarcinoma. Loss of CUL3 might be driving tumorigenesis by activating the Nrf2/RhoA pathway.

Circulating cell-free DNA as a diagnostic and prognostic biomarker for non-small-cell lung cancer: a systematic review and meta-analysis.

Aim: A meta-analysis was conducted to assess the application of circulating cell-free DNA (cfDNA) in non-small-cell lung carcinoma (NSCLC) screening, EGFR and KRAS mutation detection. Materials & methods: A comprehensive literature search was conducted. The summary sensitivity and specificity for cfDNA in NSCLC diagnosis, EGFR and KRAS mutation detection were calculated. Results: The sensitivity and specificity for NSCLC diagnosis, EGFR and KRAS mutation detection were 0.80 (95% CI: 0.72-0.87) and 0.81 (95% CI: 0.68-0.91), 0.780 (95% CI: 0.711-0.853) and 0.962 (95% CI: 0.942-0.984), 0.628 (95% CI: 0.244-0.919) and 0.959 (95% CI: 0.932-0.998), respectively. Conclusion: cfDNA was a minimally invasive approach for NSCLC diagnosis, but its clinical utility warranted more future investigations because of the suboptimal sensitivity.

Trophinin-associated protein expression is an independent prognostic biomarker in lung adenocarcinoma.

BACKGROUND: Lung cancer is the leading cause of cancer-related deaths worldwide, with lung adenocarcinoma (LAC) representing the most common subtype. Trophinin-associated protein (TROAP) is a cytoplasmic protein first identified to mediate the process of embryo transplantation, which has been recently found to be involved in microtubule regulation. However, limited information about the role of TROAP in LAC is available. METHODS: We evaluated the relationship of TROAP expression in LAC tissues with clinical pathologic parameters and the survival time in LAC patients based on a statistical analysis of The Cancer Genome Atlas (TCGA) lung cancer data (N=528). Differences in survival between high and low expression groups (median expression cutoff) from the Cox univariate/multivariate regression analysis were then compared. RESULTS: According to the Chi-square tests, we found high TROAP expression correlated with younger age (≤60) (P=0.047), male sex (P<0.005), an earlier T-stage (P=0.011), N-stage (P=0.017), M-stage (P=0.022), TNM (P=0.007), and a longer smoking history (>30 pack-year) (P<0.001). A Kaplan-Meier analysis demonstrated that high TROAP expression may correspond with poor overall survival of LAC patients in T3 stage (P=0.0013), N0 stage (P=0.014), and M0 stage (P=0.0023). Multivariate analysis confirmed that TROAP expression was related to overall survival in LAC patients independently [hazard ratio (HR): 1.784, 95% confidence interval (CI): 1.072-2.968, P=0.026]. CONCLUSIONS: Our results suggested that TROAP is an independent prognostic biomarker of poor survival in LAC.

Yap-Hippo promotes A549 lung cancer cell death via modulating MIEF1-related mitochondrial stress and activating JNK pathway.

Although the role of Yes-associated protein (Yap) has been described in the progression of lung cancer, the downstream effector of the Yap-Hippo pathway has not been identified. Accordingly, the aim of our study is to explore whether Yap modulates the activity of lung cancer by controlling mitochondrial elongation factor 1 (MIEF1)-related mitochondrial stress in a manner dependent on the JNK pathway. Cell viability was determined via MTT, LDH release and immunofluorescence assays. ATP production, the mitochondrial membrane potential, and caspase-9 activity were investigated to assess mitochondrial function. siRNA transfection and pathway blockers were used to observe the roles of MIEF1 and JNK in Yap-modulated cell viability in lung cancer cells in vitro. Yap deletion reduced cell viability in A549 and H358 lung cancer cells. At the molecular level, Yap deletion promoted mitochondrial dysfunction, as evidenced by the decreased mitochondrial potential, increased mitochondrial oxidative stress, augmented mitochondrial pro-apoptotic factor leakage and elevated caspase-9 activity. In addition, we found that Yap modulated mitochondrial stress via MIEF1 and that loss of MIEF1 abolished the regulatory actions of Yap on mitochondrial stress and cell viability. Besides, we provided evidence to support the necessary role of JNK in Yap-mediated MIEF1 upregulation. Inhibition of JNK abolished the promotive effect of Yap deletion on MIEF1 activation. Taken together, our results identified the JNK-MIEF1 pathway and mitochondrial stress as downstream effectors of Yap in lung cancer. This finding suggests a novel approach for the treatment of lung cancer in clinical practice.

CircRNA-ENO1 promoted glycolysis and tumor progression in lung adenocarcinoma through upregulating its host gene ENO1.

Lung adenocarcinoma (LUAD) has long been one of the predominant reasons for the global cancer-linked mortality. The tumor progression is shown by several studies to be promoted by increased glycolysis. Enolase 1 (ENO1), as a glycolysis enzyme, performs pivotal role in glucose metabolism and contributes to tumor progression of numerous cancers. Circular RNAs (circRNAs) are catching increasing attentions for their surging roles in regulating gene expression in cancers. Our work is to uncover the regulatory mechanism circ-ENO1 on its host gene ENO1 and its function in glycolysis and tumor progression. Circ-ENO1 and its host gene ENO1 were identified to be upregulated in LUAD cells. Functionally, silencing circ-ENO1 retarded glycolysis, inhibited proliferation, migration and EMT, induced apoptosis. The cytoplasmic localization of circ-ENO1 was determined by FISH and subcellular fractionation. Mechanistically, circ-ENO1 acted as a ceRNA to interact with miR-22-3p and upregulate ENO1 expression. In vivo experiments certified that circ-ENO1 drove tumor growth and metastasis in vivo. In summary, current study elucidated that circ-ENO1 promoted glycolysis and tumor progression in LUAD by miR-22-3p/ENO1 axis, indicating circ-ENO1 as a promising treatment target for LUAD patients.

Overexpression of APC11 predicts worse survival in lung adenocarcinoma.

BACKGROUND: Anaphase-promoting complex subunit 11 (APC11) plays an important role in gathering E2 and catalyzing ubiquitin-chain formation to support ubiquitination of substrates by acting as a catalytic core subunit of anaphase-promoting complex (APC/C). However, whether APC11 is implicated in the tumorigenesis of lung cancer is never known. MATERIALS AND METHODS: In this study, we used an online survival analysis software to estimate the prognostic value of APC11 mRNA expression level for lung cancer. Cell Counting Kit-8 assay, colony-forming assay, and transwell assay were used to assess the biological functions of APC11 in lung cancer cells. Then, 107 lung cancer patient tissues were collected to examine the expression level of APC11 by immunohistochemistry staining. Kaplan-Meier method and univariate Cox regression analysis were performed to reveal the prognostic value of APC11 protein expression in lung cancer. RESULTS: Higher mRNA level of APC11 was significantly associated with worse survival for lung adenocarcinoma, but not for lung squamous cell carcinoma. Knockdown of APC11 by siRNA apparently inhibited cell proliferation and colon formation in both H1299 and H358 cells. In addition, silencing of APC11 decreased cell migrative and invasive abilities. Moreover, immunohistochemical analysis showed that APC11 was highly expressed in lung cancer tissues, and multivariate analysis suggested that APC11 overexpression was an independent prognostic factor in lung adenocarcinoma. CONCLUSION: We suggest that APC11 could serve as a prognostic biomarker and a novel target in treating lung adenocarcinoma.

[Muscle-sparing thoracotomy in chest surgery].

OBJECTIVE: To review the clinical experience of muscle-sparing thoracotomy in intrathoracic surgery. METHODS: Thoracotomy was performed in 386 patients from 1998 to 2002, during the procedure lateral-transverse incision, free dissection of muscular flap and entering to the thoracic cavity through certain intercostal space were applied. Two sets of rib retractors were used to ensure the excellent field exposure. RESULTS: Intrathoracic surgery was carried out by this method with the advantage of excellent surgical field exposure, less pain and relative quick recovery. CONCLUSION: Muscle-sparing thoracotomy has the merits of less injury and the same good exposure as routine thoracotomy and it can be carried out in majority of chest surgery.

Optimization of lymph node dissection with VATS right upper lobectomy.

Video-assisted thoracoscopic lobectomy (VATSL) has developed rapidly. As per the requirements of tumor staging and oncotherapy, the technique of VATS mediastinal lymph node dissection plays an important role. With our experience, lymph node dissection can be performed very well on the condition that surgical process and skills be optimized.