METTL14-mediated m6A mRNA modification of G6PD promotes lung adenocarcinoma.

METTL14 functions as an RNA methyltransferase involved in m6A modification, influencing mRNA biogenesis, decay, and translation processes. However, the specific mechanism by which METTL14 regulates glucose-6-phosphate dehydrogenase (G6PD) to promote the progression of lung adenocarcinoma (LUAD) is not well understood. Quantitative measurement and immunohistochemistry (IHC) analysis have demonstrated higher levels of m6A in LUAD tissues compared to adjacent normal tissues. Additionally, the expression of METTL14 was significantly increased in LUAD tissues. In LUAD cell lines, both METTL14 and m6A levels were elevated compared to normal human lung epithelial cells. Knockdown of METTL14 markedly reduced LUAD cell proliferation, migration, and invasion. Conversely, overexpression of METTL14, but not the mutant form, significantly enhanced these cellular processes in LUAD. In vivo studies using nude mice with subcutaneously transplanted LUAD cells demonstrated that stable METTL14 knockdown led to notably reduced tumor volume and weight, along with fewer Ki67-positive cells and lung metastatic sites. Importantly, METTL14 knockdown reduced glycolytic activity in LUAD cells. Through a combination of RNA sequencing and MeRIP-sequencing, we identified numerous altered genes and confirmed that IGF2BP2 enhances G6PD mRNA stability after METTL14-mediated m6A modification, thereby promoting tumor growth and metastasis. Moreover, LUAD patients with higher levels of G6PD had poorer overall survival (OS). In conclusion, our study indicates that METTL14 upregulates G6PD expression post-transcriptionally through an m6A-IGF2BP2-dependent mechanism, thereby stabilizing G6PD mRNA. These findings propose potential diagnostic biomarkers and effective targets for anti-metabolism therapy in LUAD.

m6A modification of CDC5L promotes lung adenocarcinoma progression through transcriptionally regulating WNT7B expression.

Cell division cycle 5-like (CDC5L) protein is implicated in the development of various cancers. However, its role in the progression of lung adenocarcinoma (LUAD) remains uncertain. Our findings revealed frequent upregulation of CDC5L in LUAD, which correlated with poorer overall survival rates and advanced clinical stages. In vitro experiments demonstrated that CDC5L overexpression stimulated the proliferation, migration, and invasion of LUAD cells, whereas CDC5L knockdown exerted suppressive effects on these cellular processes. Furthermore, silencing CDC5L significantly inhibited tumor growth and metastasis in a xenograft mouse model. Mechanistically, CDC5L activates the Wnt/ß-catenin signaling pathway by transcriptionally regulating WNT7B, thereby promoting LUAD progression. Besides, METTL14-mediated m6A modification contributed to CDC5L upregulation in an IGF2BP2-dependent manner. Collectively, our study uncovers a novel molecular mechanism by which the m6A-induced CDC5L functions as an oncogene in LUAD by activating the Wnt/ß-catenin pathway through transcriptional regulation of WNT7B, suggesting that CDC5L may serve as a promising prognostic marker and therapeutic target for LUAD.

Methylprednisolone accelerate chest computed tomography absorption in COVID-19: A three-centered retrospective case control study from China.

BACKGROUND: Based on the results of some large randomized controlled trials (RCTs) confirmed the efficacy of corticosteroids in coronavirus disease 2019 (COVID-19), corticosteroids have been included in World Health Organization guidelines, but remain controversial. AIM: To investigate the efficacy and safety of low-to-moderate dose (30 to 40 mg/d) short-term methylprednisolone for COVID-19 patients. METHODS: The clinical data of 70 patients diagnosed with COVID-19 who received antiviral therapy with Arbidol for 7-10 d before admission but had no obvious absorption on chest computed tomography (CT) imaging were retrospectively analyzed. Arbidol (as the control group) and methylprednisolone (as the corticosteroid group) were given respectively after admission. After treatment, chest CT was reexamined to evaluate the absorption of pulmonary lesions. Additionally, we evaluated and compared the lymphocyte count, erythrocyte sedimentation rate (ESR), interleukin-6(IL-6), serum ferritin, lactate dehydrogenase (LDH), creatine kinase-MB (CK-MB), hypersensitive C-reactive protein (hs-CRP) and D-dimer levels, and also analyzed the incidence of toxic and side effects. RESULTS: All patients in the corticosteroid group had varying degrees of CT absorption, which was significantly better than that in the control group (CT obvious absorption rate: 89.47% vs 12.5%, P < 0.05). The average daily dose and course of methylprednisolone in the patients with significant improvement on chest CT was (38.55 ± 13.17) mg and (6.44 ± 1.86) d respectively. During the treatment, the lymphocyte count, ESR, IL-6, serum ferritin, LDH, CK-MB, hs-CRP and D-dimer levels all improved gradually, indicating that both Arbidol and methylprednisolone therapy were contributed to improving the condition of COVID-19 patients. The corticosteroid regimen did not prolong the clearance time of severe acute respiratory syndrome coronavirus 2. There were no severe adverse reactions such as gastrointestinal bleeding, secondary severe infection, hypertension, diabetic ketoacidosis, mental disorders or electrolyte disorders during the whole corticosteroid treatment process. CONCLUSION: Low-to-moderate dose short-term methylprednisolone can accelerate the chest CT imaging absorption of COVID-19 so as to improve symptoms and alleviate the condition in a short term, reduce the hospital stay, meanwhile avoid severe COVID-19 phases. The protocol has been proven to be effective and safe in clinical use.

Structure and function encoding of a bidirectional activatable synergetic DNA machine for speeded and ultrasensitive determination of microRNAs.

This work describes the unique design of a bidirectional activatable synergetic DNA machine (BAS-DNA machine) for speeded and ultrasensitive determination of microRNA-21 (miR-21), a well-known biomarker for biomedical research and early diagnosis of lung cancer. The BAS-DNA machine is composed by a pair of track strands (Track 1 and Track 2) encoding with two regions in the opposite direction for miR-21 recognition. Introduction of miR-21 can hybridize either with Track 1 or with Track 2 to activate the BAS-DNA machine with a synergistic effect for speeded amplifying the fluorescence signal. Moreover, compared with common DNA machine with only one switch for exogenous target recognition, the BAS-DNA machine with two switches for miR-21 binding allows the speeded and strong operation of the autonomous strand scission, replication, and displacement on Track 1 and Track 2 simultaneously. This behavior makes the BAS-DNA machine powerful for ultrasensitive, specific, and fast screening of miR-21 even from real biological samples, and the fluorescence signal was found to be linear from 1 pM to 10 nM with a detection limit of 703.6 fM. We envision this BAS-DNA machine with its superior assay performance will provide a new avenue for simple, sensitive, and affordable biomedical assays.

TTN/TP53 mutation might act as the predictor for chemotherapy response in lung adenocarcinoma and lung squamous carcinoma patients.

BACKGROUND: Chemotherapy is the preferred treatment in many types of cancer including lung cancer. However, most of patients resist chemotherapy resulting in disease progressive and recurrence. Titin (TTN) mutation is proved as a beneficial role in lung squamous carcinoma (LUSC), but the predictive role on chemotherapy resistance of lung cancer is still limited and discussable. METHODS: Clinical information and related somatic mutation profiles were obtained from The Cancer Genome Atlas (TCGA) database and analyzed by R-Studio using R-package. Overall survival (OS) curve and the association between gene mutation and clinical features were determined by GraphPad 6.0 software. RESULTS: Available data including 563 lung adenocarcinoma (LUAD) and 505 LUSC subjects were included in this study. Among all patients, 205 out of 563 LUAD and 326 out of 505 LUSC patients displayed TTN gene mutation. When comparing the clinical features in TTN-mutated patients to patients without TTN mutation who received chemotherapy, the tumors were always located in the upper lung in LUAD patients with TTN mutation and most of TTN-mutated subjects were at low pathological stage, which was not observed in LUSC patients. However, patients with TTN-mutation, particularly missense mutation, had a higher chemosensitivity and longer OS period than that patients without TTN mutation in both LUAD and LUSC. Of note, LUAD and LUSC patients possessed favorable OS and better chemotherapy response benefiting from TTN/tumor protein 53 (TP53) double mutation compared to TTN and TP53 mutation alone, respectively. Additionally, TTN/TP53 double mutation-initiated high rate of chemotherapy response were largely concentrated within LUAD and LUSC patients whose anatomic neoplasm subdivision were located in the upper lung. CONCLUSIONS: Collectively, TTN/TP53 co-mutation is possibly served as an effective predictor for OS and chemotherapy response in lung cancer.

IL­21/IL­21R inhibit tumor growth and invasion in non­small cell lung cancer cells via suppressing Wnt/ß­catenin signaling and PD­L1 expression.

Lung cancer is considered to be one of the world's deadliest diseases, with non­small cell lung cancer (NSCLC) accounting for 85% of all lung cancer cases. The present study aimed to investigate the role and underlying mechanisms of interleukin­21 (IL­21), and its receptor IL­21R, in NSCLC. Lung tissues and blood samples of NSCLC were used to measure IL­21, IL­21R and programmed death 1 ligand 1 (PD­L1) expression using ELISA, western blot and immunohistochemistry analyses. Following treatment with different doses of IL­21, the proliferation, invasion and migration of human NSCLC cell line A549 was evaluated using a cell counting kit­8, colony formation, Transwell and scratch wound healing assays, respectively. Additionally, IL­21R and PD­L1 expression in A549 cells was detected using western blot analysis and immunofluorescence. IL­21R silencing was subsequently used to investigate its effects in cell proliferation, invasion and migration. PD­L1, IL­1ß and tumor necrosis factor α (TNF­α) expression were measured. Finally, Wnt/ß­catenin signaling expression was evaluated using western blot analysis following treatment with IL­21. Cells were then treated with lithium chloride (LiCl), which is an agonist of Wnt/ß­catenin signaling, and the levels of PD­L1, IL­1ß and TNF­α were detected. The results revealed that IL­21 and IL­21R expression in the lung tissues and blood samples of patients with NSCLC were decreased, while PD­L1 expression was increased, compared with normal tissues or healthy controls. Treatment of A549 cells with IL­21 upregulated IL­21R expression, downregulated PD­L1 and inhibited cell growth and metastasis in a dose­dependent manner. Following IL­21R silencing, the effects of IL­21 treatment were reversed, suggesting that IL­21 acted on A549 cells through binding to IL­21R. In addition, the results demonstrated that IL­21 treatment reduced the expression levels of proteins associated with the Wnt/ß­catenin signaling, whereas activation of Wnt/ß­catenin signaling with the LiCl agonist upregulated PD­L1, IL­1ß and TNF­α expression. In conclusion, the IL­21/IL­21R axis reduced the growth and invasion of NSCLC cells via inhibiting Wnt/ß­catenin signaling and PD­L1 expression. The present results may provide a novel molecular target for NSCLC diagnosis and therapy.