Inhibition of HIST1H2AB suppresses the proliferation of lung adenocarcinoma.

BACKGROUND: Lung adenocarcinoma is one of the common causes of cancer-related deaths worldwide. Histone cluster 1 H2A family member b (HIST1H2AB) is a member of the histone H2A family. Bioinformatic analyses have revealed that HIST1H2AB is highly expressed in some cancers and might be an oncogene. However, information on the function of HIST1H2AB in lung adenocarcinoma is limited. METHODS: The expression of HIST1H2AB was analyzed in normal lung, lung adenocarcinoma and paracancerous tissues from The Cancer Genome Atlas (TCGA) database and immunohistochemistry staining. It was further verified in the relative cell lines using real-time quantitative polymerase chain reaction (RT-qPCR). When the adenocarcinoma cells lines (A549 and H1299) were successfully transfected with shHIST1H2AB or an empty plasmid packaged into a lentivirus, cell proliferation was detected using Celigo fluorescence cell-counting, colony formation and annexin V-allophycocyanin assays. Twenty nude mice were subcutaneously injected with A549 cells transfected with shHIST1H2AB or empty plasmid; the tumor size was recorded on day 25 and then measured every 3 days thereafter. The final tumor weight was measured on day 37. Significantly differentially expressed genes were analyzed using a human gene expression array. Furthermore, the potentially relevant genes were verified using RT-qPCR and western blotting. RESULTS: HIST1H2AB was highly expressed in lung adenocarcinoma tissues from TCGA database and immunohistochemistry staining. Similar results were seen in the lung adenocarcinoma cells. When the cells were successfully transfected with shHIST1H2AB or an empty plasmid, downregulation of HIST1H2AB inhibited the growth and promoted the apoptosis of lung adenocarcinoma cells. The xenograft results suggested that HIST1H2AB downregulation delayed tumor growth and reduced tumor weight. Moreover, interferon signaling pathway and four genes (HMGB1, FOXM1, F2RL1 and SLC4A7) might be regulated by HIST1H2AB in the development of lung adenocarcinoma as indicated through gene expression array, RT-qPCR and western blotting analyses. CONCLUSIONS: HIST1H2AB acts as an oncogenic protein and HIST1H2AB inhibition suppresses the proliferation of lung adenocarcinoma cells. It may be a novel target for lung adenocarcinoma therapy.

Profiles of autophagy-related genes in esophageal adenocarcinoma.

BACKGROUND: Several studies have demonstrated autophagy was involved in the process of esophageal adenocarcinoma (EAC). The aim of this study was to explore autophagy-related genes (ARGs) correlated with overall survival (OS) in EAC patients. METHODS: Expressions of ARGs in EAC and normal samples were downloaded from TCGA database. GO and KEGG enrichment analyses were used to investigate the ARGs bioinformatics functions. Univariate and multivariate cox regressions were performed to identify prognostic ARGs and the independent risk factors. ROC curve was established to evaluate the feasibility to predict the prognosis. Finally, the correlations between ARGs and clinical features were further explored. In addition, significantly different ARGs were verified in EAC specimens and normal esophageal mucosal tissues. RESULTS: Thirty significantly different ARGs were selected from EAC and normal tissues. Functional enrichments showed these ARGs were mainly related apoptosis. Multivariate cox regression analyses demonstrated eight ARGs were significantly associated with OS. Among these eight genes, BECN1 (HR = 0.321, P = 0.046), DAPK1 (HR = 0.636, P = 0.025) and CAPN1 (HR = 0.395, P = 0.004) played protective roles in survival. Gender (HR = 0.225, P = 0.032), stage (HR = 5.841, P = 0.008) and risk score (HR = 1.131, P < 0.001) were independent prognostic risk factors. ROC curves showed better efficacy to predict survival using the risk score. Additionally, we found BECN1, DAPK1, VAMP7 and SIRT1 genes were correlated significantly with survival status, gender, primary tumor and tumor stage (all P < 0.05). The experimental results confirmed the BIRC5 was overexpressed and the ITPR1, PRKN were downregulated in the EAC tissues compared with the normal esophageal mucosal tissues (all P < 0.05). CONCLUSION: Our findings suggested that autophagy was involved in the process of EAC. Several ARGs probably could serve as diagnostic and prognostic biomarkers and may help facilitate therapeutic targets in EAC patients.

The functional study of human umbilical cord mesenchymal stem cells harbouring angiotensin-converting enzyme 2 in rat acute lung ischemia-reperfusion injury model.

Acute lung ischemia-reperfusion injury (ALIRI) is featured as non-specific alveolar damage, lung edema and hypoxemia, often occurring within 72 h after surgery. It is the leading cause for primary graft failure and mortality after lung surgery and transplantation. Here we aimed to find a more effective therapeutic approach to treat ALIRI. We evaluated the combinational effects of human umbilical cord mesenchymal stem cells (HUMSCs) and angiotensin-converting enzyme 2 (ACE2) in the rat ALIRI model. HUMSCs were isolated for lentiviral-ACE2 transfection. Fifty rats were randomly divided into five groups: sham surgery, physiological saline (PS), ACE2, HUMSCs and HUMSCs-ACE2 group. Several physiological, biochemical and histological indicators were examined and compared among the five groups, such as blood oxygen saturation (Sat O2 %) and right ventricular systolic blood pressure (RVSBP), pulmonary morphology observations, several kinds of cell markers and the abundance of glutathione reductase (GR), glutathione peroxidase (GPX) and NAD(P)H quinone oxidoreductase (NQO1). Compared with HUMSCs and ACE2 groups, HUMSCs-ACE2 group showed lighter lung injuries, higher CD31 and vWF expression (endothelial cell surface markers), lower γ-H2AX (DNA damage marker) and CD68 (inflammatory cell marker) and higher anti-oxidants expression (GR, GPX and NQO1). The results indicated that HUMSCs harbouring ACE2 were more effective than either HUMSCs or ACE2 alone in alleviating the ALIRI damages. The synergistic effects of HUMSCs and ACE2 provide informative clues for mechanism study and therapeutic method development of ALIRI.

Toosendanin Induces Lung Squamous Cell Carcinoma Cell Apoptosis and Inhibits Tumor Progression via the BNIP3/AMPK Signaling Pathway.

Lung squamous cell carcinoma (LUSC) is the second most common type of non-small cell lung cancer. Toosendanin can target critical cancer cell survival and proliferation. However, the function of toosendanin in LUSC is limited. Cancer cell proliferative capacity is detected using cell morphology, colony formation, and flow cytometry. The invasiveness of the cells is detected by a Transwell assay, western blotting, and RT-qPCR. Nude mice are injected with H226 (1×106) and received an intraperitoneal injection of toosendanin every 2 days for 21 days. RNA sequence transcriptome analysis is performed on toosendanin-treated cells to identify target genes and signaling pathways. With increasing concentrations of toosendanin, the rate of cell proliferation decreases and apoptotic cells increases. The number of migrated cells significantly reduces and epithelial-mesenchymal transition is reversed. Injection of toosendanin in nude mice leads to a reduction in tumor volume, weight, and the number of metastatic tumors. Furthermore, KEGG shows that genes related to the AMPK pathway are highly enriched. BNIP3 is the most differentially expressed gene, and its expression along with phosphorylated-AMPK significantly increases in toosendanin-treated cells. Toosendanin exerts anticancer effects, induces apoptosis in LUSC cells, and inhibits tumor progression via the BNIP3/AMPK signaling pathway.

SIRT5 desuccinylates and activates SOD1 to eliminate ROS.

Cu/Zn superoxide dismutase (SOD1) is a key antioxidant enzyme. Deficiency of SOD1 is associated with various human diseases, including cancer. Here, we report that SOD1 is succinylated and that succinylation decreases its activity. SIRT5 binds to, desuccinylates and activates SOD1. SOD1-mediated ROS reduction is increased when SIRT5 is co-expressed. Furthermore, mutation of the SOD1 succinylation site inhibits the growth of lung tumor cells. These results reveal a novel post-translational regulation of SOD1 by means of succinylation and SIRT5-dependent desuccinylation, which is important for the growth of lung tumor cells.

DNAJC12 activated by HNF1A enhances aerobic glycolysis and drug resistance in non-small cell lung cancer.

Background: The DnaJ heat shock protein family member C12 (DNAJC12) gene has been demonstrated to promote lung cancer cell growth and migration by increasing ß­catenin expression. In this study, we aimed to reveal the role of DNAJC12 in modulating aerobic glycolysis and cisplatin (DDP) resistance in non-small cell lung cancer (NSCLC). Methods: Bioinformatics analysis was applied to assess the expression levels of DNAJC12 and hepatocyte nuclear factor 1-alpha (HNF1A) in NSCLC tissues and the correlation between DNAJC12 and HNF1A expression levels. Cell Counting Kit-8 (CCK-8), flow cytometry assay, and in vivo tumor formation were used to evaluate cell growth, apoptosis, and tumorigenesis. Luciferase gene reporter assay was adopted to assess the relationship between HNF1A and DNAJC12. Results: Both DNAJC12 and HNF1A were overexpressed in NSCLC tissues and cells, and their expressions showed a positive correlation. The HNF1A gene could bind to the promoter of DNAJC12 and promoted its transcription and translation. Overexpression of both DNAJC12 and HNF1A promoted cell growth, aerobic glycolysis, and inhibited cell apoptosis in NCI-H1975 and NCI-H1650 cells, as well as the cell apoptosis induced by DDP. In addition, cell growth and aerobic glycolysis mediated DNAJC12 were reversed by the silencing of ß-catenin, and downregulation of DNAJC12 abolished the above roles of HNF1A. Conclusions: This study revealed that DNAJC12, activated by the transcription factor HNF1A, could enhance aerobic glycolysis and drug resistance to DDP in NSCLC through regulating ß-catenin expression.

Phenotypic Changes of LncRNA Hotair in Non-Small-Cell Lung Cancer and Its Clinical Application.

Non-small-cell lung cancer (NSCLC) is one of the main causes of death of malignant tumors of the respiratory system. At present, the clinical demand for biomarkers for predicting and diagnosing the disease is increasing. Overexpression of LncRNA Hotair (Homeobox transcriptional antisense intergenic RNA) has been previously reported to be associated with poor prognosis and high mortality in different malignancies. qRT-PCR results showed that the expression of LncRNA Hotair in tumor tissue and serum of patients with non-small-cell lung cancer was significantly upregulated. Clinicopathological correlation analysis showed that the upregulation of LncRNA Hotair expression was closely related to lymph node metastasis and tumor lymph node metastasis (TNM) stage (P < 0.05). The results showed that transfection of pcDNA3.1-Hotair could promote the expression of LncRNA Hotair in NSCLC, while transfection of Si-Hotair could reduce the expression level of LncRNA Hotair, hinder the migration and invasion of cancer cells, and promote cell apoptosis. After transfection of Si-Hotair, molecular markers related to migration, the level of E-cadherin and Bax, increased and the level of vimentin, Bcl-2, MMP-3, VEGF, Ki-67 and PCNA decreased. This shows that the proliferation and migration of A549 cells are promoted and LncRNA Hotair deletion can inhibit the proliferation and migration of lung cancer cells. These results show that the expression level of LncRNA Hotair of NSCLC cell lines can promote the invasion and migration of NSCLC, and its expression has a significant correlation with Lymph node metastasis, tumor size, and TNM stage. Therefore, this target is of great significance for the clinical diagnosis and treatment of NSCLC.

LncRNA TMPO-AS1 facilitates the proliferation and metastasis of NSCLC cells by up-regulating ERBB2 via sponging miR-204-3p.

INTRODUCTION: This study aims at probing into the expression and biological function of long non-coding RNA (lncRNA) TMPO-AS1 in non-small cell lung cancer (NSCLC), and exploring its regulatory role for miR-204-3p and erb-b2 receptor tyrosine kinase 2 (ERBB2). METHODS: In this study, paired NSCLC samples were collected, and the expression levels of TMPO-AS1, miR-204-3p and ERBB2 were examined by quantitative real-time polymerase chain reaction (qRT-PCR); proliferative ability and colony formation ability were detected by CCK-8 assay and plate colony formation assay, respectively; flow cytometry was performed to detect the effect of TMPO-AS1 on apoptosis; Transwell assay was used to detect the changes of migration and invasion; qRT-PCR and Western blot were utilised to analyse the changes of miR-204-3p and ERBB2 regulated by TMPO-AS1; luciferase reporter gene assay and RNA immunoprecipitation assay were employed to determine the regulatory relationship between TMPO-AS1 and miR-204-3p. RESULTS: We demonstrated that TMPO-AS1 was significantly up-regulated in cancerous tissues of NSCLC samples, and positively correlated with the expression of ERBB2, while negatively correlated with miR-204-3p. After transfection of TMPO-AS1 shRNAs into NSCLC cells, the malignant phenotypes of NSCLC cells were significantly inhibited, while overexpression of TMPO-AS1 had opposite effects; TMPO-AS1 was also demonstrated to regulate the expression of miR-204-3p by sponging it, and indirectly modulate the expression of ERBB2. CONCLUSION: Collectively, we conclude that TMPO-AS1 has the potential to be the 'ceRNA' to regulate the expression of ERBB2 by sponging miR-204-3p in NSCLC.

Upregulation of microRNA-450 inhibits the progression of lung cancer in vitro and in vivo by targeting interferon regulatory factor 2.

MicroRNAs (miRNAs) are a class of non­coding RNAs that play pivotal roles in human lung cancer development. The majority of studies have focused on either non-small cell lung cancer (NSCLC) or small cell lung cancer (SCLC). In the present study, we investigated a plausible mechanism of action of miR­450 in these types of lung cancer. We found that the level of miR­450 was decreased in lung cancer cell lines, as well as in solid tumors. As exemplified in the H510A (SCLC) and H2291 (NSCLC) cells, transfection with lentivirus carrying miR­450 upregulated miR­450 expression and significantly attenuated lung cancer cell proliferation and invasion, as well as the growth of implantated tumors. Interferon regulatory factor 2 (IRF2) was also verified to be a direct target of miR­450 in lung cancer cells. The overexpression of IRF2 in the H510A and H2291 cells abrogated the inhibitory effects of miR­450 on lung cancer cell proliferation and invasion. Taken together, in this study, we identified a novel role of miR­450 in lung cancer. miR-450 targets IRF2 and thus supresses lung cancer cell proliferation and invasion.

Right native lung pneumonectomy due to over inflation three years after left single lung transplantation for pulmonary lymphangioleiomyomatosis.

Native lung hyperinflation (NLH) is one of the known complications after single lung transplantation (SLT). Generally, satisfactory results are achieved in patients undergoing SLT when simultaneous (or second stage) volume reduction of the contralateral native lung is performed. Contralateral native lung pneumonectomy after SLT is rarely reported. In this article, we report a case of a successful, right pneumonectomy of the native lung, 3 years after a left single lung transplant for pulmonary lymphangioleiomyomatosis (PLAM). The patient's pulmonary function and quality of life improved significantly after a right pneumonectomy of the native lung.

The use of CT scan and stereo lithography apparatus technologies in a canine individualized rib prosthesis.

OBJECTIVE: To design and fabricate canine rib prosthesis with full geometric shape using computed tomography (CT) scan combined with computer-aided design (CAD) and stereo lithographic (SLA) technologies and to evaluate the accuracy of this method. METHODS: After scanned on 64 rows helical CT, the cortex part of the right 7th rib was selected as the prototype for design and manufacture of the rib prosthesis and image data were stored as DICOM format. Three-dimensional (3D) surface reconstruction was applied to produce 3D image of the 7th rib and results were outputted as STL format which were then modified by UG software for establishment of CAD model. RESULTS: The rib prosthesis with full geometric shape was obtained based on CT scanning and SLA technique. About 30,000 point cloud data were acquired after 3D laser scan of the ribs. When comparing the rib prosthesis with the rib prototype, the maximum positive deviation, maximum negative deviation, average deviation and standard deviation were 1.764 mm, -2.126 mm, 0.183/-0.253 mm and 0.346 mm, respectively. There were about 88.17% of the point cloud data within the range of ±0.5 mm. CONCLUSION: It is feasible to design and fabricate rib prosthesis with full geometric shape by using CT scanning technology combined with CAD and SLA technologies. This method is fast, convenient and precise for manufacturing prosthesis. Optimization and improvement could be processed based on the deviation suggested by the scanning.

A rare case of circumferential intramural dissection of thoracic esophagus with contained esophageal perforation.

In this report, a full account of an extremely rare case on esophageal intramural dissection (EID) is presented. A 56-year-old female patient, misdiagnosed as esophageal mediastinal fistula under endoscopic view, was diagnosed correctly as EID with contained esophageal perforation in the operation and cured by thoracic esophagectomy.