Metformin inhibits human non-small cell lung cancer by regulating AMPK-CEBPB-PDL1 signaling pathway.

Metformin has been found to have inhibitory effects on a variety of tumors. However, its effects on non-small cell lung cancer (NSCLC) remain unclear. We demonstrated that metformin could inhibit the proliferation of A549 and H1299 cells. RNA transcriptome sequencing revealed that PDL1 was significantly downregulated in both cell types following treatment with metformin (P < 0.001). Jaspar analysis and chromatin immunoprecipitation showed that CEBPB could directly bind the promoter region of PDL1. Western blotting showed that protein expression of the isoforms CEBPB-LAP*, CEBPB-LAP, and CEBPB-LIP was significantly upregulated and the LIP/LAP ratio was increased. Gene chip analysis showed that PDL1 was significantly upregulated in A549-CEBPB-LAP cells and significantly downregulated in A549-CEBPB-LIP cells (P < 0.05) compared with CEBPB-NC cells. Dual-luciferase reporter gene assay showed that CEBPB-LAP overexpression could promote transcription of PDL1 and CEBPB-LIP overexpression could inhibit the process. Functional assays showed that the changes in CEBPB isoforms affected the function of NSCLC cells. Western blotting showed that metformin could regulate the function of NSCLC cells via AMPK-CEBPB-PDL1 signaling. Animal experiments showed that tumor growth was significantly inhibited by metformin, and atezolizumab and metformin had a synergistic effect on tumor growth. A total of 1247 patients were retrospectively analyzed, including 166 and 1081 patients in metformin and control groups, respectively. The positive rate of PDL1 was lower than that of the control group (HR = 0.338, 95% CI = 0.235-0.487; P < 0.001). In conclusion, metformin inhibited the proliferation of NSCLC cells and played an anti-tumor role in an AMPK-CEBPB-PDL1 signaling-dependent manner.

Identification and validation of an immune cell infiltrating score predicting survival in patients with lung adenocarcinoma.

BACKGROUND: Immune infiltration may predict survival and have clinical significance in lung cancer. However, immune signatures derived from immune profiling based on bulk tumor transcriptomes have not been systematically established in lung adenocarcinoma. We aimed to construct an immune cell infiltrating score, using a new algorithm for evaluating immune infiltration, to improve the prognostic model of lung adenocarcinoma. METHODS: Public datasets of lung adenocarcinoma from the Gene Expression Omnibus and The Cancer Genome Atlas were adopted as the training and validation cohorts. Fractions of different immune cell subtypes in each sample were estimated using the CIBERSORT algorithm. The immune infiltrating score was further developed by a least absolute shrinkage and selection operator regression model. The prognostic value and clinical relationship of the model was then further explored. RESULTS: An immune infiltrating score model was established on the basis of the immune cells in the training cohort. A high score was associated with significantly worse survival in patients with lung adenocarcinoma (P < 0.001). The prognostic value of the score was confirmed in the validation cohort. The immune infiltrating score could improve the accuracy of predictions of survival when combined with the staging system. Furthermore, the score was potentially associated with patient smoking status and histologic subtype of lung adenocarcinoma. Its possible association with the efficacy of adjuvant chemotherapy was not statistically significant. CONCLUSION: The immune cell infiltrating score has prognostic significance in predicting overall survival in patients with lung adenocarcinoma.

Non-lung cancer specific mortality after lobectomy or sublobectomy in patients with stage IA non-small cell lung cancer ≤2 cm: A propensity score analysis.

BACKGROUND AND OBJECTIVES: To investigate non-lung cancer specific mortality between stage IA non-small cell lung cancer (NSCLC) tumors less than and equal to 2 cm treated with lobectomy and sublobectomy. METHODS: Surveillance, epidemiology, and end results database was queried for patients who underwent lobectomy and sublobectomy. Propensity score matching (PSM) was used to achieve balance in clinicopathological characteristics. We used Fine-and-Gray hazard functions to analyze cause-specific mortality and risk factors. Standardized mortality ratios were calculated to describe cause specific mortality relative to the general population. RESULTS: After PSM, 3,844 patients underwent lobectomy and 1,922 patients underwent sublobectomy. Three leading causes of non-lung cancer mortality were cardiovascular disease (CVD), chronic obstructive pulmonary diseases (COPD), and other cancers. The 5-year cumulative non-lung cancer mortality of lobectomy and sublobectomy groups were 11.4% and 14.0%, respectively (P = .090). Multivariate analyses revealed that age, sex, histology, tumor size, and marital status (P < .01) were independent predictors of non-lung cancer specific mortality. In both groups, risks of CVD specific mortality were comparable to that in the general population, whereas the risk of COPD specific mortality was higher relative to the general population. CONCLUSIONS: As a significant competing event, non-lung cancer specific mortality is comparable between stage IA NSCLC tumors less than equal to 2 cm treated with lobectomy and sublobectomy.

[Analysis of the effect of different pathological subtypes to prognosis in stage I pulmonary adenocarcinoma].

OBJECTIVE: To analyze the prognostic value of the new classification (proposed by International Association for the Study of Lung Cancer/American Thoracic Society/European Respiratory Society) in stage I pulmonary adenocarcinoma. METHODS: Pathological slides of 328 cases of stage I pulmonary invasive adenocarcinoma were reviewed according to the new classification. The patients received operation in Department of Thoracic Surgery of Zhongshan Hospital affiliated to Fudan University from January 2005 to December 2009. There were 145 male and 183 female patients with an average age of (59 ± 10) years (ranging from 34 to 82 years). Clinical, pathological, and survival data were retrospectively analyzed. Kaplan-Meier method was used for analysis of survival, and Cox regression analysis was used for finding out prognostic factors. RESULTS: Five-year progression-free survival rate and overall survival rate of lepidic-predominant subtype were both 100%. Five-year progression-free survival rate of patients with micropapillary component (49.3%) was significantly lower than that of patients without micropapillary component (75.4%, χ² = 8.154, P = 0.004). Regression analysis showed that tumor size is an independent prognostic factor of death (HR = 1.967, 95% CI: 1.507 to 2.567, P = 0.000) and recurrence (HR = 1.796, 95% CI: 1.469 to 2.198, P = 0.000). In subgroup analysis, the presence of solid component (HR = 1.985, 95% CI: 1.013 to 3.888, P = 0.046) and tumor size (HR = 1.941, 95% CI: 1.455 to 2.589, P = 0.000) were independent prognostic factors of recurrence for stage IB pulmonary adenocarcinoma. CONCLUSIONS: The new classification of adenocarcinoma is of prognostic value in stage I pulmonary adenocarcinoma. The presence of solid or micropapillary component impacts on survival. Detailed record of each component in tumor is necessary.

Identification of reference miRNAs in human tumors by TCGA miRNA-seq data.

Although the accuracy of detecting the expression of miRNAs by quantitative real-time polymerase chain reaction (qRT-PCR) is highly dependent on reliable reference miRNAs, many commonly used reference miRNAs are not stably expressed and as such are not suitable for quantification and normalization of qRT-PCR data. To solve this problem, we analyzed the global expression profiles of thousands of samples in 14 types of common human tumors released by The Cancer Genome Atlas (TCGA), and identified the most stably and highly expressed miRNAs as candidate reference miRNAs in each type of tumor. We found that miR-361-5p and let-7i-5p were the most recommended candidate reference miRNAs in nine and eight types of tumors, respectively, followed by let-7a-5p, mir-28-5p and miR-99b-5p. Our results are of important value to those researchers focused on miRNA; however, these candidate reference miRNAs still need to be validated prior to their use in qRT-PCR studies.

Clinical outcome of subcentimeter non-small cell lung cancer after surgical resection: single institution experience of 105 patients.

BACKGROUND: The detection of subcentimeter lung cancers has significantly improved with advances in computed tomography and the emergence of screening protocols. We reviewed the clinicopathological features and surgical outcomes of patients with subcentimeter non-small cell lung cancer (NSCLC) in our institution. METHODS: A total of 105 patients who underwent lobectomy or sublobar resection for subcentimeter NSCLCs were retrospectively reviewed. Clinicopathological characteristics and survival were analyzed statistically using Student's t-test for continuous variables, Fisher's exact for categorical variables and Cox regression for multivariable analysis. RESULTS: A total of 105 patients (35 male, 70 female; mean age 61.4, range 38-77 years) were analyzed. Patients underwent lobectomy (n = 71), segmentectomy (n = 19), or wedge resection (n = 15). The overall 5-year survival was 91.3%. No significant differences were observed in overall and recurrence-free survival after segmentectomy or lobectomy; patients undergoing wedge resection had shorter survival compared to those who underwent lobectomy. Elevated preoperative serum CEA levels and positive nodal status correlated with poorer survival, and were identified as independent prognostic factors in multivariate analysis. CONCLUSION: Systematic nodal dissection is recommended for patients with subcentimeter NSCLC. Segmentectomy offers comparable oncologic results to lobectomy. Elevated preoperative serum CEA level implies shorter survival for patients with these tiny tumors.

Transcription factor ZNF263 enhances EGFR-targeted therapeutic response and reduces residual disease in lung adenocarcinoma.

EGF receptor (EGFR) tyrosine kinase inhibitors (TKIs) have achieved clinical success in lung adenocarcinoma (LUAD). However, tumors often show profound but transient initial response and then gain resistance. We identify transcription factor ZNF263 as being significantly decreased in osimertinib-resistant or drug-tolerant persister LUAD cells and clinical residual tumors. ZNF263 overexpression improves the initial response of cells and delays the formation of persister cells with osimertinib treatment. We further show that ZNF263 binds and recruits DNMT1 to the EGFR gene promoter, suppressing EGFR transcription with DNA hypermethylation. ZNF263 interacts with nuclear EGFR, impairing the EGFR-STAT5 interaction to enhance AURKA expression. Overexpressing ZNF263 also makes tumor cells with wild-type EGFR expression or refractory EGFR mutations more susceptible to EGFR inhibition. More importantly, lentivirus or adeno-associated virus (AAV)-mediated ZNF263 overexpression synergistically suppresses tumor growth and regrowth with osimertinib treatment in xenograft animal models. These findings suggest that enhancing ZNF263 may achieve complete response in LUAD with EGFR-targeted therapies.

Selective mediastinal lymphadenectomy without intraoperative frozen section examinations for clinical stage I non-small-cell lung cancer: retrospective study of 403 cases.

OBJECTIVE: The extent of mediastinal lymphadenectomy for clinical stage I non-small-cell lung cancer (NSCLC) remains controversial. This study explored the value of selective mediastinal lymphadenectomy from the clinical viewpoint. METHODS: From 2005 to 2008, a total of 403 patients diagnosed clinically as having stage I NSCLC underwent lobectomy and mediastinal lymph node dissection. Among them, 309 underwent complete mediastinal lymphadenectomy, and the other 94 underwent selective mediastinal lymphadenectomy. We compared the perioperative parameters and overall survival statistics for the two groups retrospectively. RESULTS: The two groups had no significant differences in sex, pathology, tumor location, or preoperative staging. The selective mediastinal lymphadenectomy group had an older average age, with a much higher rate of patients >70 years of age (p = 0.016). Also, the patients were apt to undergo thoracoscopic lobectomy (p = 0.044). This group had shorter operating times and less intraoperative bleeding. No significant differences in total drainage volume, length of hospital stay, or complication rates were found between the two groups. The mean follow-up periods were 35.8 ± 13.7 vs. 34.6 ± 17.2 months. Local and distant recurrence rates were 25.6 % vs. 30.9 %, respectively (p = 0.560). The 3-year and 5-year overall survival rates were 83.0 % and 74.6 % vs. 75.1 % and 68.5 %, respectively (p = 0.216). CONCLUSIONS: For patients with clinical stage I NSCLC, selective mediastinal lymphadenectomy can reduce the trauma caused by the procedure, especially for elderly patients and those with co-morbidities. Survival was acceptable and was no worse than that after complete mediastinal lymphadenectomy. Our results need to be confirmed by prospective randomized controlled studies.

Impact of a Ground-glass Opacity Component on c-Stage IA Lung Adenocarcinoma.

Studies have shown that tumors with ground-glass opacity (GGO) components are associated with favorable outcomes. However, this view should be confirmed in an international cohort. We aimed to verify the impact of a GGO component on clinical (c)-stage IA lung adenocarcinoma and to describe the biological discrepancies between the part-solid and pure-solid groups. We evaluated 1333 cases of surgically resected c-stage IA lung adenocarcinomas, including 484 part-solid and 849 pure-solid tumors. Furthermore, we matched the solid size between the 2 groups and examined 470 patients. We compared the prognoses between the 2 groups before and after matching. The prognostic and biological differences were described before and after matching. Compared with the pure-solid group, the part-solid group was associated with favorable outcomes [5-year overall survival (OS) 99.4% vs 87.6%, P < 0.001; 5-year recurrence-free survival (RFS) 96.9% vs 82.2%, P < 0.001]. Similar results were obtained after matching (5-year OS 98.9% vs 92.2%, P = 0.012; 5-year RFS 95.0% vs 88.5%, P = 0.007). Multivariable analyses revealed that GGO component appearance was a factor of better OS and RFS. The part-solid tumor, regardless of the size of the solid component, had a similar outcome to the pure-solid tumor of c-stage T1a classification. Also, more epidermal growth factor receptor, human epidermal growth factor receptor-2 mutations, and receptor tyrosine kinase ROS-1-positive were observed in the part-solid group. In comparison, more wild types and Kirsten-Ras were observed in the pure-solid group. Adenocarcinomas with a GGO component were associated with superior outcomes. The GGO component should be considereda new clinical T descriptor. Early-stage lung adenocarcinomas with and without a GGO component may be 2 distinct tumor types.

Ground glass nodules with scattered or eccentric island-shaped consolidations may have poor outcomes.

Background: To explore the effect of scattered or eccentric shaped types of ground glass opacity (GGO) on outcomes of patients with solid-dominant peripheral lung adenocarcinoma. Methods: We evaluated patients with solid-dominant peripheral lung adenocarcinoma, who underwent radical surgery at Zhongshan Hospital, Fudan University, between January 2013 and December 2015. Morphologically heterogeneous solid-dominant lung adenocarcinoma in imaging findings was based on the last preoperative computed tomography (CT) scans. Endpoints were recurrence-free survival (RFS) and overall survival (OS). Kaplan-Meier analysis and the log-rank test were used to estimate survival differences. Impact factors were assessed by univariable logistic regression analysis. Results: We retrospectively collected data from 200 patients, including 170 patients with central island-shaped CT imaging, 18 patients with scattered shaped CT imaging, and 12 patients with eccentric shaped CT imaging. Eleven patients experienced recurrence or metastases. Kaplan-Meier survival curves showed significant survival differences between the central island-shaped type and scattered shaped or eccentric shaped type for OS (c-stage IA: 5-year OS: 100% vs. 92.1%; HR = 0.019, p = 0.0025; p-stage IA: 100% vs. 95.2%; HR = 0.146, p = 0.1139) and RFS (c-stage IA: 5-year RFS: 100% vs. 59.7%; HR = 0.001, p < 0.0001; p-stage IA: 100% vs. 64.5%; HR < 0.001, p < 0.0001). Univariable logistic regression analysis showed that scattered and eccentric shaped types were related to poor outcomes (p < 0.012, odds ratio = 18.8). Conclusions: Relative spatial position of GGO and solid components may affect patient outcomes. Patients with scattered or eccentric shaped GGO may have a poor prognosis.

Expression and clinical significance of UCH37 in human esophageal squamous cell carcinoma.

BACKGROUND: Ubiquitin carboxyl-terminal hydrolase 37 (UCH37), a member of the DUBs, was found to play an important role in oncogenesis through promoting some Proto-oncogenes' expression and stem cell-like characteristics in the cell in previous research. The aim of this study was to assess the value of UCH37 in predicting tumor recurrence after curative resection in esophageal squamous cell carcinoma (ESCC) patients. METHODS: We analyzed UCH37 protein expression in 111 clinicopathologically characterized ESCC cases, from those who underwent curative resection between 2007 and 2008, by immunohistochemistry. The prognostic significance was assessed using Kaplan-Meier survival estimates and log-rank tests. RESULTS: We found that UCH37 expression was higher in the cancer tissue than in non-tumorous control tissue at protein level and was overexpressed in tumor tissues of recurrent patients. There was a significant difference of UCH37 expression in patients categorized according to TNM stage (p = 0.038) and lymph nodes metastasis condition (p = 0.009). Univariate analyses revealed that UCH37 was a significant predictor for overall survival and disease-free survival, and multivariate analyses showed that UCH37 was an independent prognostic marker for ESCC recurrence. A prognostic significance of UCH37 was also found in the subgroup of lymph nodes metastasis condition classification. About 90 % of the recurrent patients recurred within 2 years, of which 84.4 % were predicted by UCH37. CONCLUSION: UCH37 is associated with outcome and recurrence of ESCC and can be a novel predictor for poor prognosis of ESCC patients after curative resection.

Primary pulmonary mucoepidermoid carcinoma: an analysis of 21 cases.

BACKGROUND: The optimal treatment for pulmonary mucoepidermoid carcinoma (MEC), a rare type of tumor, has not been established yet. This study analyzed the survival of pulmonary MEC patients and attempted to find clues for optimal treatment. METHODS: A total of 21 patients with pulmonary MEC from November 2004 to January 2011 were included in the investigation. Immunohistochemistry, epidermal growth factor receptor (EGFR) mutation, and survival were retrospectively studied. RESULTS: Among the 21 pulmonary MEC patients, 17 were diagnosed with low-grade malignancy and 4 with high-grade malignancy through pathological examination. The prognosis was found to be poor in the presence of lymph nodes. The expression rates of EGFR and HER2 were 28.6% and 0%, respectively, which correlated with neither grade nor prognosis. The mutation rate of EGFR was 0. Log-rank test results indicated that age, grade, lymph node metastasis, and tumor-node-metastasis stage were prognostic factors. CONCLUSION: Age, grade, lymph node metastasis and tumor-node-metastasis stage correlate with the survival of pulmonary MEC patients. TRIAL REGISTRATION: This study was approved and registered by the Ethics Committee of Zhongshan Hospital. Written informed consent was obtained from all participants prior to treatment.

Pan-Driver-Negatives versus Epidermal Growth Factor Receptor Mutants for C-Stage IA Lung Adenocarcinoma with Ground-Glass Opacity.

PURPOSE: We aimed to verify the prognosis of epidermal growth factor receptor (EGFR) mutation of clinical (c)-stage IA lung adenocarcinoma with the ground-glass opacity (GGO) component. METHODS: We evaluated 226 cases of surgically resected c-stage IA lung adenocarcinoma with GGO component. Endpoints were overall survival (OS) and recurrence-free survival (RFS). Kaplan-Meier analysis and the log-rank test were used to estimate the survival differences. Prognostic factors were assessed using the univariable and multivariable Cox proportional hazards model. RESULTS: Among the 226 cases, 177 cases harbored the EGFR-mutant adenocarcinoma with the GGO component. The mean duration of follow-up time was 54.4 ± 1.2 months. The 5-year OS and RFS did not differ significantly between the EGFR-mutant and wild-type groups (5-year OS 100% vs. 94.3%, hazard ratio [HR] 0.276, P = 0.168; 5-year RFS 94.7% vs. 95.7%, HR 0.873, P = 0.864). Multivariable Cox hazard model revealed that radiologically solid component size (P = 0.010) and pathological node-positive (P = 0.036) were significant predictors of an inferior RFS. CONCLUSION: EGFR-mutant was not a prognostic factor of OS and RFS for c-stage IA lung adenocarcinoma with the GGO component. Radiologically solid component size and pathological lymph node status were independent prognostic factors of worse RFS.

Expression Analysis of Ligand-Receptor Pairs Identifies Cell-to-Cell Crosstalk between Macrophages and Tumor Cells in Lung Adenocarcinoma.

Background: Lung adenocarcinoma is one of the most commonly diagnosed malignancies worldwide. Macrophage plays crucial roles in the tumor microenvironment, but its autocrine network and communications with tumor cell are still unclear. Methods: We acquired single-cell RNA sequencing (scRNA-seq) (n = 30) and bulk RNA sequencing (n = 1480) samples of lung adenocarcinoma patients from previous literatures and publicly available databases. Various cell subtypes were identified, including macrophages. Differentially expressed ligand-receptor gene pairs were obtained to explore cell-to-cell communications between macrophages and tumor cells. Furthermore, a machine-learning predictive model based on ligand-receptor interactions was built and validated. Results: A total of 159,219 single cells (18,248 tumor cells and 29,520 macrophages) were selected in this study. We identified significantly correlated autocrine ligand-receptor gene pairs in tumor cells and macrophages, respectively. Furthermore, we explored the cell-to-cell communications between macrophages and tumor cells and detected significantly correlated ligand-receptor signaling pairs. We determined that some of the hub gene pairs were associated with patient prognosis and constructed a machine-learning model based on the intercellular interaction network. Conclusion: We revealed significant cell-to-cell communications (both autocrine and paracrine network) within macrophages and tumor cells in lung adenocarcinoma. Hub genes with prognostic significance in the network were also identified.

FSCN1 induced PTPRF-dependent tumor microenvironment inflammatory reprogramming promotes lung adenocarcinoma progression via regulating macrophagic glycolysis.

PURPOSE: Macrophages (MΦs) play a dual role in the promotion and suppression of lung adenocarcinoma (LUAD), the function of which is influenced by the metabolic status. The role of protein tyrosine phosphatase receptor type F (PTPRF) in cancer has not been elucidated, and its role in MΦs remains to be seen. METHODS: The Seahorse XFe 96 Cell Flow Analyzer detected glucose metabolism in tumor cells and macrophages. The expressions of FSCN1, M-CSF, IL4, PTPRF and IGF1 in macrophages were detected by Western blotting and qRT-PCR. Binding of FSCN1 and IGF1R was detected by co-immunoprecipitation. The tumor status in animals was observed using the IVIS Lumina III imaging system. RESULTS: We found that Fascin Actin-Bundling Protein 1 (FSCN1) activates the PI3K-AKT and JAK-STAT signaling pathways in LUAD cells via binding to IGF-1R, thereby promoting the secretion of cytokines such as IL4 and M-CSF. IL4 and M-CSF promote the expression of PTPRF in MΦs, leading to M2 polarization of MΦs by increasing glucose intake and lactate production. In return, M2-type MΦs act on LUAD cells by secreting cytokines such as IGF-1, CCL2, and IL10, which ultimately promote tumor progression. In vivo experiments proved that the knockdown of FSCN1 in A549 cells and PTPRF in MΦs greatly reduced LUAD proliferative and metastatic capacity, which was consistent with the in vitro findings. CONCLUSIONS: This study investigated the reprogramming effects of FSCN1 and PTPRF on inflammatory cytokines in the LUAD microenvironment, revealing potential mechanisms by which FSCN1 and PTPRF promote tumor progression and providing a new experimental basis for LUAD treatment.

Solid Components in the Mediastinal Window of Computed Tomography Define a Distinct Subtype of Subsolid Nodules in Clinical Stage I Lung Cancers.

BACKGROUND: We aimed to validate the clinicopathologic characteristics and prognostic value of the presence of solid components in the mediastinal window of computed tomography scan in clinical stage I pulmonary subsolid nodules (SSNs). METHODS: We retrospectively evaluated patients with pulmonary SSNs resected between 2011 and 2016. We classified SSNs into heterogeneous ground-glass nodules (HGGNs) (solid component detected only in lung window) and part-solid nodules (PSNs) (solid component detected both in lung/mediastinal windows). RESULTS: A total of 487 patients (216 PSNs) were included. PSNs were associated with higher frequencies of micropapillary or solid pathologic patterns (18.1% vs. 3.3%; P < .001), epidermal growth factor receptor gene mutation (39.4% vs. 32.8%), and other types of gene mutations (2.3% vs. 1.1%; P = .043). Logistic regression analysis revealed that male sex (odds ratio [OR], 2.58; 95% confidence interval [CI], 1.20-5.57; P = .016) and higher consolidation tumor ratio (CTR) (OR, 110.04; 95% CI, 8.56-1414.39; P < .001) remained independent for invasive adenocarcinomas with poor differentiation. Receiver operating characteristic analyses revealed that solid component size in the mediastinal window (area under the curve [AUC], 0.731; 95% CI, 0.653-0.808; P < .0001) showed a better predictive ability to poor differentiation compared with solid component size in the lung window and CTR. The 5-year recurrence-free survival (RFS) rate of PSNs was worse than that of HGGNs (94.6% vs. 99.1%; P = .019). Multivariate Cox regression revealed that positive lymph node status (hazard ratio, 22.99; 95% CI, 4.52-116.86; P < .001) indicated worse RFS for PSNs. CONCLUSION: SSNs with solid components in mediastinal window demonstrated clinicopathologic and prognostic features different from those without in clinical stage I lung cancer. Solid components in mediastinal window was a strong predictor of poor differentiation.

Circadian rhythm-associated clinical relevance and Tumor Microenvironment of Non-small Cell Lung Cancer.

Objective: We aimed to explore the prognostic implication for non-small cell lung cancer (NSCLC) based on the expression profiles of circadian clock-related genes (CCRGs), and describe the changes of immune infiltration and cell functions of related to the circadian rhythm. Methods: Univariate and multivariate Cox proportional hazard regression were performed to determine a CCRGs risk-score significantly correlated with overall survival (OS) of the training set and validation set. GO, KEGG, and GSVA indicated discrepant changes in cellular processes and signaling pathways associated with these CCRGs. Immune cell infiltration and mutation rates were investigated by the online analysis platform and the algorithm provided by works of literature. Results: The signature-based on ten-gene signatures could independently predict the OS both in TCGA lung adenocarcinoma (p < 0.001, HR: 1.228, 95% CI: 1.158 to 1.302) and lung squamous cell carcinoma (p < 0.001, HR: 2.501, 95% CI: 2.010 to 3.117), respectively. The circadian oscillations driven by CCRGs could disturb the metabolism and cellular functions of cancer cells. The infiltration level of critical cells in specific anti-tumor immunity process was suppressed apparently. In contrast, the infiltrating of inflammatory cells and immune cells with negative regulatory effects were promoted in the high-risk group. CCRGs were evolutionarily conserved with low mutation rates, which brought difficulties to explore therapeutic targets. Conclusion: We identified and validated a circadian rhythm signature to described clinical relevance and tumor microenvironment of NSCLC, which revealed that circadian rhythms might play an influential role in the NSCLC.

Prognostic Impact of Radiological Consolidation Tumor Ratio in Clinical Stage IA Pulmonary Ground Glass Opacities.

OBJECTIVES: Our study aimed to validate pathologic findings of ground-glass nodules (GGOs) of different consolidation tumor ratios (CTRs), and to explore whether GGOs could be stratified according to CTR with an increment of 0.25 based on its prognostic role. METHODS: We retrospectively evaluated patients with clinical stage IA GGOs who underwent curative resection between 2011 and 2016. The patients were divided into 4 groups according to CTR step by 0.25. Cumulative survival rates were calculated by the Kaplan-Meier method. Univariate and multivariate Cox regression analyses were conducted to obtain the risk factors on relapse-free survival (RFS). The surv_function of the R package survminer was used to determine the optimal cutoff value. Receiver operating characteristic (ROC) analysis was generated to validate optimal cutoff points of factors. RESULTS: A total of 862 patients (608 women; median age, 59y) were included, with 442 patients in group A (CTR ≤ 0.25), 210 patients in group B (0.25

Multi-omics characterization and validation of MSI-related molecular features across multiple malignancies.

HEADINGS AIMS: To establish a microsatellite instability (MSI) predictive model in pan-cancer and compare the multi-omics characterization of MSI-related molecular features. MATERIALS AND METHODS: We established a 15-gene signature for predicting MSI status and performed a systematic assessment of MSI-related molecular features including gene and miRNA expression, DNA methylation, and somatic mutation, in approximately 10,000 patients across 30 cancer types from The Cancer Genome Atlas, Gene Expression Omnibus database, and our institution. Then we identified common MSI-associated dysregulated molecular features across six cancers and explored their mutual interfering relationships and the drug sensitivity. KEY FINDINGS: we demonstrated the model's high prediction performance and found the samples with high-MSI were mainly distributed in six cancers: BRCA, COAD, LUAD, LIHC, STAD, and UCEC. We found RPL22L1 was up-regulated in the high-MSI group of 5/6 cancer types. CYP27A1 and RAI2 were down-regulated in 4/6 cancer types. More than 20 miRNAs and 39 DMGs were found up-regulated in MSI-H at least three cancers. We discovered some drugs, including OSI-027 and AZD8055 had a higher sensitivity in the high MSI-score group. Functional enrichment analysis revealed the correlation between MSI score and APM score, HLA score, or glycolysis score. The complicated regulatory mechanism of tumor MSI status in multiple dimensions was explored by an integrated analysis of the correlations among MSI-related genes, miRNAs, methylation, and drug response data. SIGNIFICANCE: Our pan-cancer study provides a valuable predictive model and a comprehensive atlas of tumor MSI, which may guide more precise and personalized therapeutic strategies for tumor patients.

HIF-1α switches the functionality of TGF-ß signaling via changing the partners of smads to drive glucose metabolic reprogramming in non-small cell lung cancer.

BACKGROUND: Most cancer cells have fundamentally different metabolic characteristics, particularly much higher glycolysis rates than normal tissues, which support the increased demand for biosynthesis and promote tumor progression. We found that transforming growth factor (TGF)-ß plays a dual function in regulating glycolysis and cell proliferation in non-small cell lung cancer. METHODS: We used the PET/MRI imaging system to observe the glucose metabolism of subcutaneous tumors in nude mice. Energy metabolism of non-small cell lung cancer cell lines detected by the Seahorse XFe96 cell outflow analyzer. Co-immunoprecipitation assays were used to detect the binding of Smads and HIF-1α. Western blotting and qRT-PCR were used to detect the regulatory effects of TGF-ß and HIF-1α on c-MYC, PKM1/2, and cell cycle-related genes. RESULTS: We discovered that TGF-ß could inhibit glycolysis under normoxia while significantly promoting tumor cells' glycolysis under hypoxia in vitro and in vivo. The binding of hypoxia-inducible factor (HIF)-1α to the MH2 domain of phosphorylated Smad3 switched TGF-ß function to glycolysis by changing Smad partners under hypoxia. The Smad-p107-E2F4/5 complex that initially inhibited c-Myc expression was transformed into a Smad-HIF-1α complex that promoted the expression of c-Myc. The increased expression of c-Myc promoted alternative splicing of PKM to PKM2, resulting in the metabolic reprogramming of tumor cells. In addition, the TGF-ß/Smad signal lost its effect on cell cycle regulatory protein p15/p21. Furthermore, high expression of c-Myc inhibited p15/p21 and promoted the proliferation of tumor cells under hypoxia. CONCLUSIONS: Our results indicated that HIF-1α functions as a critical factor in the dual role of TGF-ß in tumor cells, and may be used as a biomarker or therapeutic target for TGF-ß mediated cancer progression.