Analysis of the Expression and Clinical Significance of VIPR1 in Lung Adenocarcinoma Based on TCGA Database

Background: The expression of VIPR1 is associated with the prognosis of many malignant tumors. To analyze the expression and clinical significance of vasoactive intestinal peptide Types I (VIPR1) in lung adenocarcinoma based on The Cancer Genome Atlas (TCGA)database. Methods and results：The RNASeq data, clinical data and prognosis data of lung adenocarcinoma and normal lung tissue were downloaded from TCGA database. The expression difference of VIPR1 mRNA in lung adenocarcinoma and normal lung tissue（p<0.05）, and the correlations of the expression levels of VIPR1 in lung adenocarcinoma with clinical pathological characteristics and prognosis were analyzed. The possible regulatory signaling pathways of VIPR1 were predicted by the gene set enrichment analysis (GSEA). CIBERSORT was used to analyze the expression of immune cells in tumor tissues and normal tissues. TIMER was used to analyze the relationship between VIPR1 and immune cell expression. The expression levels of VIPR1 mRNA in lung adenocarcinoma tissues were significantly lower than that in normal lung tissues. The expression levels of VIPR1 mRNA were significantly correlated with gender, T staging，N staging and pathological grade（p<0.05）but not with age, M staging and survival status. Survival analysis showed that the survival time of patients with low expression of VIPR1 was significantly lower than that of patients with high expression. In addition, the expression level of VIPR1 in lung adenocarcinoma was negatively correlated with the infiltration level of myeloid inhibitory cells (r = -0.448, p < 0.01). Through GSEA functional enrichment analysis, it was found that VIPR1 was mainly related to cell cycle pathway, p53 signal pathway, DNA replication pathway, RNA degradation pathway and so on.Conclusions：VIPR1 gene may be a new target for the clinical prognosis and targeted therapy of lung adenocarcinoma.

LncRNA OGFRP1 acts as an oncogene in NSCLC via miR-4640-5p/eIF5A axis

Background Long noncoding RNAs (lncRNAs) OGFRP1 is up-regulated in endometrial cancer and cervical carcinoma, and OGFRP1 suppression inhibits the malignant behavior of cancer cells. Here, we evaluated the expression pattern, biological function and potential mechanism of OGFRP1 in non-small cell lung cancer (NSCLC). Methods The expression of target genes in 25 pairs of clinically collected NSCLC and normal lung tissue samples was detected by qRT-PCR or western blot. We screened the siRNA (siOGFRP1) to down-regulate the expression of OGFRP1 in A549 and H1299 cells. The biological function of A549 and H1299 cells were examined by CCK8, wound healing and transwell assays. The molecular mechanism of OGFRP1 was further explored. Results The expression of OGFRP1 in NSCLC tissues were higher than that in normal lung tissue. siOGFRP1 inhibited the proliferation, migration and invasion of A549 and H1299 cells. In addition, the expression of EMT-related and apoptosis-related proteins was changed by siOGFRP1 transfection. OGFRP1 can directly interact with miR-4640-5p, and siOGFRP1 increased the level of miR-4640-5p. Moreover, miR-4640-5p could directly bind to the 3’ UTR region of eIF5A mRNA. eIF5A was highly expressed in NSCLC tissues, and predicted a poor prognosis. In addition, the expression of miR-4640-5p and eIF5A in NSCLC tissues were negatively correlated, while the expression of OGFRP1 and eIF5A were positively correlated. Knockdown of OGFRP1 inhibited the expression of eIF5A, while transfection of miR-4640-5p inhibitor up-regulated the expression of eIF5A. Conclusions Taken together, we demonstrated that down-regulation of OGFRP1 inhibited the progression of NSCLC through miR-4640-5p/eIF5A axis.

The effect of constant light and melatonin on urethane-induced lung carcinogenesis in mice and expression of clock genes and proteins

Introduction. Circadian rhythms are an adaptational mechanism to day-night cycle. At the cellular level rhythms are supported by oscillations of transcription of Bmal1, 2, Clock and Npas2, Per1-3, Cry1, 2 clock genes; in the organism rhythms are synchronized with melatonin, the "hormone of darkness". Reliable data have been obtained that disruption of biological "clocks" at the cellular and/or organism level are associated with carcinogenesis, but the experimental results in this direction remain incomplete. Aim. Assessment of the continuous lighting and melatonin administration effects on chemically induced carcinogenesis in mice and expression of clock genes and proteins in normal and tumor lung tissue. Materials and methods. Lung tumors were induced by urethane in 120 male SHR mice. Animals were kept at constant (LL) or standard (LD) light; half of the animals received melatonin (MT) daily at 20 mg/l at night with water for 45 weeks of experiment. At the end of the experiment, the frequency and multiplicity of lung tumours of different size were assessed. Expression of clock genes Clock, Bmal1, Cry1 was assessed in tumor samples and normal lung tissue by real-time PCR; protein content of BMAL1, CLOCK, CRY1 and PER2 was determined by immunohistochemistry. Statistical data were processed using the programs MS Excel 2007, GraphPad Prism 6.0 with commonly used statistical criteria. Results. The number of animals with lung tumors was between 89 and 100% in different groups. Large tumors (>2 mm) were more frequently observed in LL group (62 tumors out of 294, 21,1%) than in LD group (59 tumors out of 405, 14,6%, p=0,0245). Administration of exogenous melatonin at constant lighting statistically significantly reduced the frequency of large tumors (28 tumors out of 320, 8.8%, p=0,0001 in comparison with LL). In normal lung tissue of animals kept at constant lighting (LL) the increase in relative expression of clock genes in comparison with LD group was revealed: Bmal1 by 3,1 times (p=0,02) and Cry1 by 3,6 times (p=0,0002). No such differences were found for Clock gene. The relative content of BMAL1 and CLOCK proteins in all experimental conditions was higher in adenomas and adenocarcinomas compared to normal tissue. Conclusion. Constant lighting promotes the development of chemically induced lung tumors, and melatonin administration inhibits carcinogenesis under constant lighting. The content of BMAL1 and CLOCK transcription activator proteins in lung tumors was found to increase in comparison with normal tissue, whereas no increase in the expression level of corresponding genes in tumors was observed.

Metformin Protects against Radiation-Induced Acute Effects by Limiting Senescence of Bronchial-Epithelial Cells

Radiation-induced damage to normal lung parenchyma remains a dose-limiting factor in thorax-associated radiotherapy (RT). Severe early and late complications with lungs can increase the risk of morbidity in cancer patients after RT. Herein, senescence of lung epithelial cells following RT-induced cellular stress, or more precisely the respective altered secretory profile, the senescence-associated secretory phenotype (SASP), was suggested as a central process for the initiation and progression of pneumonitis and pulmonary fibrosis. We previously reported that abrogation of certain aspects of the secretome of senescent lung cells, in particular, signaling inhibition of the SASP-factor Ccl2/Mcp1 mediated radioprotection especially by limiting endothelial dysfunction. Here, we investigated the therapeutic potential of a combined metformin treatment to protect normal lung tissue from RT-induced senescence and associated lung injury using a preclinical mouse model of radiation-induced pneumopathy. Metformin treatment efficiently limited RT-induced senescence and SASP expression levels, thereby limiting vascular dysfunctions, namely increased vascular permeability associated with increased extravasation of circulating immune and tumor cells early after irradiation (acute effects). Complementary in vitro studies using normal lung epithelial cell lines confirmed the senescence-limiting effect of metformin following RT finally resulting in radioprotection, while fostering RT-induced cellular stress of cultured malignant epithelial cells accounting for radiosensitization. The radioprotective action of metformin for normal lung tissue without simultaneous protection or preferable radiosensitization of tumor tissue might increase tumor control probabilities and survival because higher radiation doses could be used.

The effect of miR-496 on eIF3h in lung invasive adenocarcinoma

This study explores the correlation between microRNA-496 (miR-496) expression and eukaryotic translation initiation factor 3H (eIF3h) expression in lung invasive adenocarcinoma. A total of 71 patients with lung adenocarcinoma were included in the study. The tumor tissue samples were collected together with the normal lung tissue samples that were acquired more than 2 cm away from the edge of the tumor). Expression of miR-496 was detected using real-time fluorescence quantitative PCR. Expression of Ki67 and Bcl-2 in lung adenocarcinoma was detected with Western blotting, and expression of eIF3hwas detected with immunohistochemistry. Expression of miR-496 was significantly lower in the tumor than in normal lung tissue (P<0.05). miR-496 expression was significantly decreased in the tumor samples from the patients with large tumor maximum diameter, pleural recidivism, lymph node metastasis, and TNM stage III + IV, compared to the samples from the patients with small tumor diameter, no pleural recidivism, no lymph node metastasis, and TNM stage I + II (P<0.05). No significant difference in miR-496 expression was identified among the groups of different sex, age, and tumor differentiation degree (P>0.05). Low expression of miR-496 correlated with a shorter survival time of the patients (P<0.05). Moreover, miR-496 expression was negatively correlated with Ki67, Bcl-2 and eIF3h expression. We speculate that and miR-496 has the potential to regulates eIF3h expression, and low expression of miR-496 in lung adenocarcinoma may be related to the occurrence and development of the tumor. Mir-496 may be an independent prognostic factor in lung adenocarcinoma.

Intratumoural heterogeneity and immune modulation in lung adenocarcinoma of female smokers and never smokers

Lung cancer is still the leading cause of cancer death worldwide despite declining smoking prevalence in industrialised countries. Although lung cancer is highly associated with smoking status, a significant proportion of lung cancer cases develop in patients who never smoked, with an observable bias towards female never smokers. A better understanding of lung cancer heterogeneity and immune system involvement during tumour evolution and progression in never smokers is therefore highly warranted. We employed single nucleus transcriptomics of surgical lung adenocarcinoma (LADC) and normal lung tissue samples from patients with or without smoking history. Immune cells as well as fibroblasts and endothelial cells respond to tobacco smoke exposure by inducing a highly inflammatory state in normal lung tissue. In the presence of LADC, we identified differentially expressed transcriptional programmes in macrophages and cancer-associated fibroblasts, providing insight into how the niche favours tumour progression. Within tumours, we distinguished eight subpopulations of neoplastic cells in female smokers and never smokers. Through pseudotemporal ordering, we inferred a trajectory towards two differentiated tumour cell states implicated in cancer progression and invasiveness. A proliferating cell population sustaining tumour growth exhibits differential immune modulating signatures in both patient groups. Our results resolve cellular heterogeneity and immune interactions in LADC, with a special emphasis on female never smokers and implications for the design of therapeutic approaches.

A Study of The Histopathological Effect of White Albino Mice Lung Infected with Aspergillus Fumigatus, Treated with Amphotericin B, And Trametes Spp.

The present study was designed to investigate the histological lesions in the lungs of white mice infected with Aspergillus fumigatus by intraperitoneal injection, nasal implantation, the current research also examined an attempt to treat these tissue lesions by using both of the antifungal Amphotericin B and the alcoholic extract of Trametes for two weeks. The results showed that A. fumigatus led to tissue changes represented by the degeneration, desquamation the lining respiratory epithelium of the bronchiole, thickening, rupturing of the alveolar walls, congestion of the vessels and lymphocytic infiltration, the lungs of mice that infected with A. fumigatus and treated with the antifungal Amphotericin B and alcoholic extract of the Trametes spp. showed improvement, as degeneration and desquamation of the lining of the respiratory epithelium of the trachea, the presence of dust cells, lymphocytic infiltration, in addition to the appearance of semi-normal lung tissue in another histological also were observed. We conclude that Amphotericin B and alcoholic extract of Trametes spp. have activity in treating the histological lesions that resulted from A. fumigatus infection.

Phospholipid dynamics in ex vivo lung cancer and normal lung explants

In cancer cells, metabolic pathways are reprogrammed to promote cell proliferation and growth. While the rewiring of central biosynthetic pathways is being extensively studied, the dynamics of phospholipids in cancer cells are still poorly understood. In our study, we sought to evaluate de novo biosynthesis of glycerophospholipids (GPLs) in ex vivo lung cancer explants and corresponding normal lung tissue from six patients by utilizing a stable isotopic labeling approach. Incorporation of fully 13C-labeled glucose into the backbone of phosphatidylethanolamine (PE), phosphatidylcholine (PC), and phosphatidylinositol (PI) was analyzed by liquid chromatography/mass spectrometry. Lung cancer tissue showed significantly elevated isotopic enrichment within the glycerol backbone of PE, normalized to its incorporation into PI, compared to that in normal lung tissue; however, the size of the PE pool normalized to the size of the PI pool was smaller in tumor tissue. These findings indicate enhanced PE turnover in lung cancer tissue. Elevated biosynthesis of PE in lung cancer tissue was supported by enhanced expression of the PE biosynthesis genes ETNK2 and EPT1 and decreased expression of the PC and PI biosynthesis genes CHPT1 and CDS2, respectively, in different subtypes of lung cancer in publicly available datasets. Our study demonstrates that incorporation of glucose-derived carbons into the glycerol backbone of GPLs can be monitored to study phospholipid dynamics in tumor explants and shows that PE turnover is elevated in lung cancer tissue compared to normal lung tissue.