ZEB1-AS1 is associated with poor prognosis in non-small-cell lung cancer and influences cell migration and apoptosis by repressing ID1.

Long non-coding RNAs (lncRNAs) have been reported to play a vital role in non-small-cell lung cancer (NSCLC). ZEB1-AS1 overexpression predicts a poor prognosis in osteosarcoma and colorectal cancers. In the current study, we determined the clinical significance and prognostic value of ZEB1-AS1 in patients with NSCLC. The expression of ZEB1-AS1 and inhibitor of differentiation-1 (ID1) was measured using qRT-PCR and Western blot. Cell growth, migration, and invasion were determined using colony formation assays, Transwell assay, and flow cytometry, respectively. Tumor growth was determined with a xenograft model. ZEB1-AS1 was significantly up-regulated in NSCLC tissues compared with normal samples. ZEB1-AS1 overexpression was significantly associated with advanced tumor, lymph node, and metastases (TNM) stage and tumor size, as well as poorer overall survival. Moreover, ZEB1-AS1 knockdown inhibited NSCLC cell proliferation and migration, and promoted cell apoptosis. In addition, a chromatin immunoprecipitation assay revealed that ZEB1-AS1 interacted with STAT3, thereby repressing ID1 expression. Furthermore, rescue experiments indicated that ZEB1-AS1 functioned as an oncogene partly by repressing ID1 in NSCLC cells. Taken together, our findings indicate that ZEB1-AS1 serves as a promising therapeutic target to predict the prognosis of NSCLC.

Elevated serum soluble programmed cell death ligand 1 concentration as a potential marker for poor prognosis in small cell lung cancer patients with chemotherapy.

BACKGROUND: Potential relationship between serum soluble programmed cell death ligand 1 and prognosis of small cell lung cancer is not well explored. The aim of the study was to reveal the prognostic significance of serum soluble programmed cell death ligand 1 in patients with small cell lung cancer. METHODS: A total of 250 small cell lung cancer patients and 250 controls were included. Research information was obtained from their medical records. Blood samples were collected on admission. Serum concentration of programmed cell death ligand 1 was measured using Enzyme-Linked Immunosorbent Assay. The patients underwent cisplatin-etoposide chemotherapy with a maximum of six cycles. Subsequently, they were followed-up for 12 months, and therapeutic response and cancer death were recorded. RESULTS: Serum concentration of programmed cell death ligand 1 was higher in the patients than in the controls on admission (P < 0.001). After chemotherapy, 112 patients had no response to this therapy. In the 12-month follow up period, 118 patients died due to this cancer. Multivariate Cox regression model revealed that the higher serum concentration of programmed cell death ligand 1 on admission was associated with the higher risk of no response to chemotherapy or cancer caused death (HR: 1.40, 95% CI: 1.05 ~ 1.87; HR: 1.43, 95% CI: 1.08 ~ 1.87). CONCLUSION: Elevated serum concentration of soluble programmed cell death ligand 1 might be an independent risk factor for non-response to chemotherapy and cancer caused death in small cell lung cancer patients.

Circular RNA circ-PLCD1 functions as a tumor suppressor in non-small cell lung cancer by inactivation of PI3K/AKT signaling pathway.

Circular RNAs (circRNAs) are emerging as crucial regulators in tumorigenesis and aggressive progression. However, their biological roles in non-small cell lung cancer (NSCLC) remain largely unknown. Here, by performing circRNA high throughput sequencing in 4 paired NSCLC and normal tissues, we found a NSCLC-associated circRNA, circ-PLCD1, which was evidently downregulated in NSCLC tissues and cell lines. Circ-PLCD1 was transcriptionally activated by tumor-inhibiting protein p53, and exogenous expression of circ-PLCD1 inhibited NSCLC cell proliferation, invasion and induced apoptosis. Mechanistically, circ-PLCD1 acted as a competitive endogenous RNA (ceRNA) to sponge miR-375 and miR-1179 and elevate PTEN, a well-known inhibitor of oncogenic PI3K/AKT signaling, thereby repressing NSCLC tumorigenesis. Importantly, we also identified this ceRNA regulatory axis of circ-PLCD1/miR-375/miR-1179/PTEN in vivo by establishing a xenograft tumor model. Clinically, NSCLC patients with low circ-PLCD1 expression had larger tumor size, later clinical stage and shorter survival time than those with high circ-PLCD1 expression. Altogether, our findings reveal the important tumor suppressive role of circ-PLCD1 in NSCLC, reactivation of this circRNA may be considered as a novel therapeutic avenue for patient with NSCLC.

Relationship between tumor mutational burden, gene mutation status, and clinical characteristics in 340 cases of lung adenocarcinoma.

Tumor mutational burden (TMB) is an emerging predictive marker of response to immune checkpoint inhibitor therapies. We evaluated the correlation between clinical indicators and high-throughput sequencing results and TMB in lung adenocarcinoma patients, with the aim of finding simpler and more economical factors as surrogate markers for TMB. The medical records, next-generation sequencing data, and immunohistochemistry results of 340 lung adenocarcinoma patients who were admitted to the First Affiliated Hospital of Zhengzhou University between 2019 and 2020 were collected. The mutated genes were screened for, and the obtained mutated genes were subjected to functional enrichment analysis using R software. A protein-protein interaction (PPI) network was also constructed, and significant modules in the network were identified. Gene Ontology (GO) analyses were performed for the core genes. Univariate and multivariate correlation analyses were performed to judge the correlation between gene mutations and TMB. Genes with a junction mutation rate >1 were selected to construct PPI network and 13 high-connection core genes were screened. The results of GO enrichment analysis showed that the biological processes related to mutant core genes mainly included mitotic cell cycle and cell aging. Subsequently, ATM (p = 0.006) and PIK3CA (p = 0.008) mutation positivity were identified by univariate and multivariate correlation analysis, while TP53 (p = 0.003) and EGFR (p = 0.008) mutation negativity were significantly associated with elevated TMB. The results of this study demonstrate that ATM- and PIK3CA-positive and EGFR-negative mutation status are strongly associated with high levels of TMB and have the potential to be predictive biomarkers of response to immune checkpoint inhibitors in lung adenocarcinoma patients.

Intracranial complete response to toripalimab and anlotinib in a patient with recurrent brain metastases of small cell lung cancer after failure of second-line maintenance therapy: a case report.

Background: Approximately 10-25% of patients with small cell lung cancer (SCLC) have brain metastases at the time of diagnosis. Radiotherapy is a common treatment for brain metastases, but the relapse rates are high. Accumulating evidence suggests that immunotherapy may have a better therapeutic effect for brain metastases. Here, we reported a patient with limited-stage SCLC and relapsed brain metastases who achieved sustained intracranial complete response (CR) to programmed cell death-1 (PD-1) inhibitor toripalimab and multikinase inhibitor anlotinib. Case Description: A 59-year-old female patient developed brain metastases after initial treatment for limited stage SCLC. CR of brain lesions was achieved after intensity-modulated radiation therapy followed by chemotherapy with irinotecan plus lobaplatin and concurrent anlotinib. PD-1 inhibitor sintilimab combined with anlotinib were given as maintenance therapy. Small and asymptomatic brain lesions relapsed 2.5 months after achieving CR. Another three cycles of sintilimab combined with anlotinib failed to control the relapsed brain lesions. Following two cycles of another PD-1 inhibitor toripalimab combined with anlotinib, the relapsed brain metastases disappeared. Then the patient received another seven cycles of this regimen with sustained CR, and no serious adverse reactions occurred. Interestingly, the primary lung tumor achieved sustained CR from the end of initial treatment to the last follow-up. Conclusions: This case suggests that toripalimab in combination with anlotinib may be a promising treatment option for patients with brain metastases from SCLC.

[Regulating action of iron regulatory protein-2 in iron metabolism of lung cancer].

OBJECTIVE: To discuss the regulating mechanism of iron regulatory protein-2 (IRP2) in the iron metabolism of lung cancer. METHODS: The cultured A549 cells were divided into 3 groups: liposome group (including liposomes 20 mg/L), random oligonucleotide group (SCODN group) and antisense oligonucleotide group (ASODN group). And the liposome-mediated transfection was employed with the liposome and SCODN groups as controls. Reverse transcription-polymerase chain reaction (RT-PCR) and Western blot were used to examine the mRNA and protein expressions of iron metabolism-related transferring (Tf), transferrin receptor (TfR) and ferritin (Fn) genes, etc. RESULTS: After a 48-hour transfection, the mRNA expression of Tf had no statistically significant difference among three groups (F = 2.18, P = 0.078); the mRNA expression of TfR in the ASODN group was significantly lower than that in the liposome and SCODN groups (P < 0.05). The expression of Fn mRNA in the ASODN group (0.56 ± 0.06) was higher than that in the liposome (0.36 ± 0.05) and SCODN groups (0.39 ± 0.03) (P < 0.05). After a 48-hour transfection, the IRP2 protein expression of the ASODN group was significantly lower than those of the liposome and SCODN groups (P < 0.05). The Tf protein expression was not statistically different in three groups (F = 2.67, P = 0.088). The TfR protein expression of the ASODN group was lower than those of the liposome and SCODN groups (P < 0.05). And the Fn protein expression of the ASODN group was higher than those of the liposome and SCODN groups (P < 0.05). CONCLUSION: IRP2 may affect the expressions of TfR and Fn in lung adenocarcinoma A549 cells by changing the amount of protein and regulating the iron metabolism.

[Levels of HMGB1 in induced sputum from patients with asthma and chronic obstructive pulmonary disease].

OBJECTIVE: To explore the relationship between the sputum levels of high mobility group protein B1 (HMGB1) and airway inflammation in bronchial asthma and chronic obstructive pulmonary disease (COPD) patients. METHODS: A total of 57 patients with persistent asthma [per Global Initiative for Asthma (GINA) guidelines], 30 patients with stable COPD [stratified by Global Initiative for COPD (GOLD) status] and 20 control subjects were recruited. After completing an asthma control questionnaire, spirometry was performed before sputum induction. The ratio of forced expiratory volume in the first second (FEV(1))/predictive value (FEV(1)%Pre) and neutrophil differential count in induced sputum were recorded. The concentrations of HMGB1 in the supernatant of sputum were measured by ELISA (enzyme-linked immunosorbent assay). RESULTS: The sputum concentrations of HMGB1 in the asthmatics and COPD patients were significantly higher than those of the control subjects [(291 ± 55) and (511 ± 39) vs (61 ± 5) ng/L, all P < 0.01]. And they were significantly negatively correlated with FEV(1)%Pre in all subjects. The levels of HMGB1 in induced sputum of COPD patients were significantly higher than those of asthmatics and healthy controls (P < 0.01). No significant difference existed in the levels of HMGB1 between patients with eosinophilic and noneosinophilic asthma [(290 ± 55) vs (292 ± 54) ng/L, P > 0.05]. The HMGB1 levels with COPD stage II and stage III were significantly higher than those with stage I [(526 ± 29) and (541 ± 29) vs (471 ± 18) ng/L]. The differences of sputum neutrophil percentage were statistically significant in mild, moderate and severe asthma [(27 ± 2)%, (36 ± 4)%, (49 ± 4)%]. And the sputum levels of HMGB1 were significantly higher in the patients with moderate and severe asthma [(312 ± 14) vs (347 ± 11) ng/L]. And the levels of HMGB1 in asthmatic and COPD patients were positively correlated with neutrophil percentage. According to the multivariate analysis, neutrophil percentage and FEV(1)%Pre were independent predictors of sputum HMGB1, but not smoking, age, gender and eosinophilic percentage. CONCLUSION: HMGB1 may contribute to airway inflammation through its higher expression in bronchial asthma and COPD patients.

[Changes of HMGB1 and RAGE in induced sputum from patients with bronchial asthma].

OBJECTIVE: To explore the relationship between HMGB1 (high mobility group box-1) protein and receptor for advanced glycosylation end products (RAGE) and the nosogenesis and severity of bronchial asthma. METHODS: Based on the criteria, the asthma group included 64 acute-onset asthma patients while the control group had 20 healthy cases. The asthma group received a 4-week combination inhalation therapy of budesonide and formoterol. Lung functions and induced sputum examinations were conducted before and after treatment. The percentage of a second forced expiratory volume in the predicted value (FEV(1)%) was recorded. A differential count of neutrophilic leukocyte in reduced sputum was performed. And the sputum levels of HMGB1 and RAGE were detected by ELISA (enzyme linked immunosorbent assay). RESULTS: Prior to treatment, the neutrophilic leukocyte percentage and the levels of HMGB1 and RAGE were all higher than those of control group (P < 0.01). The induced sputum of severe asthma patients had higher levels of neutrophilic leukocyte percentage and HMGB1 than those of mild cases (P < 0.01). But the level of RAGE showed no statistical significance among mild, moderate or severe asthma cases (P > 0.05). The post-treatment levels of neutrophilic leukocyte percentage, HMGB1 and RAGE were lower as compared with the pre-treatment ones (P < 0.01). These three parameters in uncontrolled cases were higher than those in completely controlled cases (P < 0.05); in asthma group, both HMGB1 and RAGE had a negative correlation with FEV(1)% (r = -0.830, r = -0.632, P < 0.01); in induced sputum, both HMGB1 and RAGE had a positive correlation with FEV(1)% (r = 0.820, r = 0.623, P < 0.01). The levels of HMGB1 and RAGE were positively correlated (r = 0.929, P < 0.01). CONCLUSION: Both HMGB1 and RAGE participate in the inflammatory process of asthmatic airway. HMGB1 is correlated with the severity of asthma. And the levels of HMGB1 and RAGE in induced sputum may be employed as reference indices for the observation of therapeutic effects.

Overexpression of miR-1290 contributes to cell proliferation and invasion of non small cell lung cancer by targeting interferon regulatory factor 2.

MicroRNAs are small endogenous non-coding RNAs, which can frequently emerge as regulators in many cancer types. MiR-1290 was found to be abnormally elevated in non small cell lung cancer (NSCLC). However, the underlying molecular mechanism still needs to be investigated. Here, we demonstrated that miR-1290 expression levels were remarkably upregulated in NSCLC tissues compared to adjacent normal tissues. Higher miR-1290 expression levels positively associated with lymph node metastasis and advanced tumor stage. Functional assays showed that upregulated miR-1290 expression in NSCLC cells enhanced cell proliferation, cell colony formation and invasion capacities in vitro. Furthermore, we found that miR-1290 promoted cell proliferation related protein CDK2 and CDK4 expression and enhanced Epithelial-Mesenchymal Transition (EMT) process by downregulating E-cadherin expression and upregulating N-cadherin expression. Bioinformatics analysis and luciferase reporter gene assays revealed that Interferon regulatory factor 2 (IRF2) was a direct target of miR-1290. Overexpression of miR-1290 can degrade IRF2 mRNA and downregulated IRF2 protein expression in NSCLC cells. Upregulated IRF2 could partly rescue the promoting effects induced by miR-1290 overexpression on cell proliferation and invasion of NSCLC. Additionally, we confirmed that reduced miR-1290 expression could suppress tumor growth using a tumor xenograft model in vivo. Thus, we concluded that miR-1290 may serve as a potential target of NSCLC treatment.