Cell Line IMR-90 Research Articles

Targeting SOX18 Transcription Factor Activity by Small-Molecule Inhibitor Sm4 in Non-Small Lung Cancer Cell Lines

The transcription factor SOX18 has been shown to play a crucial role in lung cancer progression and metastasis. In this study, we investigated the effect of Sm4, a SOX18 inhibitor, on cell cycle regulation in non-small cell lung cancer (NSCLC) cell lines LXF-289 and SK-MES-1, as well as normal human lung fibroblast cell line IMR-90. Our results demonstrated that Sm4 treatment induced cytotoxic effects on all three cell lines, with a greater effect observed in NSCLC adenocarcinoma cells. Sm4 treatment led to S-phase cell accumulation and upregulation of p21, a key regulator of the S-to-G2/M phase transition. While no significant changes in SOX7 or SOX17 protein expression were observed, Sm4 treatment resulted in a significant upregulation of SOX17 gene expression. Furthermore, our findings suggest a complex interplay between SOX18 and p21 in the context of lung cancer, with a positive correlation observed between SOX18 expression and p21 nuclear presence in clinical tissue samples obtained from lung cancer patients. These results suggest that Sm4 has the potential to disrupt the cell cycle and target cancer cell growth by modulating SOX18 activity and p21 expression. Further investigation is necessary to fully understand the relationship between SOX18 and p21 in lung cancer and to explore the therapeutic potential of SOX18 inhibition in lung cancer.

DIFFERENTIAL EXPRESSION AND STABILITY OF XIAP AND CIAP-1 MAY INFLUENCE THE APOPTOSIS-RESISTANT PHENOTYPE OF LUNG FIBROBLASTS IN IDIOPATHIC PULMONARY FIBROSIS

DIFFERENTIAL EXPRESSION AND STABILITY OF XIAP AND CIAP-1 MAY INFLUENCE THE APOPTOSIS-RESISTANT PHENOTYPE OF LUNG FIBROBLASTS IN IDIOPATHIC PULMONARY FIBROSIS

Bioactive Cell-Derived ECM Scaffold Forms a Unique Cellular Microenvironment for Lung Tissue Engineering.

Chronic lung diseases are one of the leading causes of death worldwide. Lung transplantation is currently the only causal therapeutic for lung diseases, which is restricted to end-stage disease and limited by low access to donor lungs. Lung tissue engineering (LTE) is a promising approach to regenerating a replacement for at least a part of the damaged lung tissue. Currently, lung regeneration is limited to a simplified local level (e.g., alveolar–capillary barrier) due to the sophisticated and complex structure and physiology of the lung. Here, we introduce an extracellular matrix (ECM)-integrated scaffold using a cellularization–decellularization–recellularization technique. This ECM-integrated scaffold was developed on our artificial co-polymeric BETA (biphasic elastic thin for air–liquid interface cell culture conditions) scaffold, which were initially populated with human lung fibroblasts (IMR90 cell line), as the main generator of ECM proteins. Due to the interconnected porous structure of the thin (<5 µm) BETA scaffold, the cells can grow on and infiltrate into the scaffold and deposit their own ECM. After a mild decellularization procedure, the ECM proteins remained on the scaffold, which now closely mimicked the cellular microenvironment of pulmonary cells more realistically than the plain artificial scaffolds. We assessed several decellularization methods and found that 20 mM NH4OH and 0.1% Triton X100 with subsequent DNase treatment completely removed the fibroblasts (from the first cellularization) and maintains collagen I and IV as the key ECM proteins on the scaffold. We also showed the repopulation of the primary fibroblast from human (without chronic lung disease (non-CLD) donors) and human bronchial epithelial (16HBE14o−) cells on the ECM-integrated BETA scaffold. With this technique, we developed a biomimetic scaffold that can mimic both the physico-mechanical properties and the native microenvironment of the lung ECM. The results indicate the potential of the presented bioactive scaffold for LTE application.

Cytotoxic and Apoptotic Effects of Chemogenic and Biogenic Nano-sulfur on Human Carcinoma Cells: A Comparative Study.

Two-dimensional nanostructures have gained tremendous interest in the field of biomedical applications and cancer activity in particular. Although sulfur is known for its wide range of biological activities, its potentiality in two-dimensional forms as an antitumor agent is hitherto unexplored. To address the current deficient knowledge on nano-sulfur as an antitumor agent, we report the synthesis of nano-sulfur sheets/particles and their cytotoxic, apoptotic activity against human carcinoma cell lines. In vitro cytotoxic effects of biogenic nanosheets (SNP-B) and chemogenic nanoparticles (SNP-C) were assessed against human lung carcinoma (A549), human epidermoid carcinoma (A431), human promyelocytic leukaemia (HL60) and human lung fibroblast (IMR90) cell lines. Cell cycle analysis, apoptotic study, and caspase-3 expression studies were carried out to understand the mechanism of cytotoxic activity of nano-sulfur. The MTT assay indicated a dose-dependent decrease in viability of all the cell lines treated with nano-sulfur, with SNP-B being more toxic compared to SNP-C. The apoptotic study and cell cycle analysis indicated cell cycle arrest followed by apoptosis-induced cell death. The caspase-3 expression study indicated that nano-sulfur induces apoptosis by the activation of caspase through the mitochondrial pathway. Apart from this, a lower cytotoxicity was observed in IMR90 cell lines treated with SNP-B , indicating a higher specificity of synthesized nanosheets towards cancer cells. Taken all together, this work highlights the potentiality of sulfur nanosheets in inducing cytotoxicity and apoptotic activity, and the impact of morphology as a critical determinant on the cytotoxic response on various cell lines.

Anlotinib Alleviates Pulmonary Fibrosis by Inhibiting PFKFB3-Driven Glycolysis via the Downregulation of PCBP3

Background: Idiopathic pulmonary fibrosis (IPF) is a fatal disease in which the normal alveolar network is gradually replaced by fibrotic scars. Current evidence implicates the differentiation of fibroblasts into myofibroblasts is correlated with metabolic alterations in IPF. Here, we tried to decipher whether anlotinib, a novel multitargeted tyrosine kinase inhibitor, could alleviate bleomycin (BLM) induced pulmonary fibrosis and explore the possible mechanisms. Methods: In this study, C57BL/6 mice were intraperitoneally injected with anlotinib immediately or 7 days after BLM injection to investigate the preventative and therapeutic effects of anlotinib on BLM-induced pulmonary fibrosis. Additionally, we used primary mouse lung fibroblasts and the IMR90 cell line to examine the effects and the mechanism of anlotinib on fibroblasts. Findings: We observed that anlotinib limited BLM-induced lung fibrosis and inhibited the lactate accumulation in mice. Meanwhile, we found that anlotinib could inhibit the PFKFB3-dependent glycolysis in fibroblasts and diminish myofibroblasts differentiation. The mechanism by which anlotinib exerts potent antifibrotic effects was by downregulating the RNA binding protein (RBP) PCBP3, decreasing PFKFB3 translation and inhibiting glycolysis in fibroblasts. Interpretation: Our data suggest that anlotinib has antifibrotic effects on the lungs and provide a possible mechanism. Anlotinib may be a novel and potent candidate for protection against pulmonary fibrosis. We also identify PCBP3 that is involved in the regulation of glycolysis reprogramming and lung fibrogenesis, which could be a potential therapeutic target for pulmonary fibrosis. Fund: This work was supported by National Natural Science Foundation of China (No. 81870058). Declaration of Interests: The authors declare that they have no conflict of interest. Ethics Approval Statement: All experiments were conducted in accordance with protocols approved by the Southern Medical University Institutional Animal Care and Use Committee.

Selina-1,3,7(11)-trien-8-one and Oxidoselina-1,3,7(11)-trien-8-one from Eugenia uniflora Leaf Essential Oil and Their Cytotoxic Effects on Human Cell Lines.

The sesquiterpenes selina-1,3,7(11)-trien-8-one and oxidoselina-1,3,7(11)-trien-8-one were isolated from the essential oil of Eugenia uniflora L. leaves. The structures were elucidated using spectrometric methods (UV, GC–MS, NMR, and specific optical rotation). The relationship between antioxidant activity, as determined by DPPH assay, and the cytotoxic effect was evaluated using tumor cells, namely lung adenocarcinoma epithelial cells (A549) and human hepatoma carcinoma cells (HepG2), as well as a model of normal human lung fibroblast cells (IMR90). Both compounds did not show prominent free-radical scavenging activity according to DPPH assay, and did not inhibit lipid peroxidation in Wistar rat brain homogenate. The isolated compounds showed pro-oxidative effects and cytotoxicity in relation to the IMR90 cell line.

Flavopiridol causes cell cycle inhibition and demonstrates anti-cancer activity in anaplastic thyroid cancer models.

Anaplastic thyroid cancer (ATC) is a rare, but nearly uniformly fatal disease that is typically resistant to chemotherapy and radiation. Alternative strategies to target this cancer at a molecular level are necessary in order to improve dismal outcomes for ATC patients. We examined the effects of flavopiridol, a CDK inhibitor, in a panel of ATC cell lines. When cell lines were treated over a ten-point concentration range, CAL62, KMH2 and BHT-101 cell lines had a sub micromolar half-maximal inhibitory concentration, while no effect was seen in the non-cancerous cell line IMR-90. Flavopiridol treatment resulted in decreased levels of the cell cycle proteins CDK9 and MCL1, and induced cell cycle arrest. Flavopiridol also decreased the in vitro ability of ATC cells to form colonies and impeded migration using a transwell migration assay. In vivo, flavopiridol decreased tumor weight and tumor volume over time in a patient-derived xenograft model of ATC. Given the observed in vitro and in vivo activity, flavopiridol warrants further investigation for treatment of ATC.

Abstract 4069: Selective ablation of solid tumors using a p53-targeted FAST-LNP gene therapy

Abstract While chemotherapy is a key treatment strategy for many solid tumors, it is rarely curative as patients will eventually become resistant. In this study, we sought to develop an effective suicide gene therapy approach for solid tumors that specifically exploits their unique transcriptional activation state. The tumor suppressor p53 is frequently mutated or dysregulated in cancer, and as a result the upstream signaling pathways activating p53 transcription are strongly upregulated. RNA-seq analysis has demonstrated that p53 transcription is significantly upregulated in almost all forms of cancer. Additionally, HCT116 cells lacking functional p53 display a 6-fold increase in p53 promoter activity when compared to its wild type p53 parent cell line. To exploit this, we have developed a Fusogenix FAST-LNP formulation to deliver a p53-driven inducible suicide gene, iCasp9, to solid tumors and destroy them upon activation with small molecule dimerizer, Rapamycin. To establish a proof-of-concept, plasmid encoding iCasp9 and luciferase under control of the p53 promoter was constructed and evaluated in a panel of cancer cell lines. While LNPs administered without Rapamycin or Rapamycin administered alone had no impact on cell viability, we observed greater than 90% apoptotic cell death when both were employed in a wide range of cancer cell lines with p53 deletions or mutations, as measured using cell viability assays, imaging assays, as well as Annexin V and TUNEL flow cytometry. Induction of iCasp9 protein expression and caspase-mediated apoptosis was confirmed using Western blot. No cell death was observed in cells with intact p53 such as human umbilical vein endothelial cells or the fibroblast cell line IMR-90. Next, we assessed the efficacy of FAST-LNPs containing p53-iCasp9 in xenograft PC-3 and H1299 models of human prostate cancer and lung cancer respectively. In some experiments, tumors were implanted subcutaneously in the flanks of 30 mice and allowed to grow to 500 mm3 before treatment by intravenous doses of 100 µg LNP twice per week during continuous low dosing of Rapamycin. We observed a rapid and dramatic reduction in tumor volume averaging 87% over the following 48 hours, with durable response. Tumors in control mice continued to grow exponentially. Overall survival of mice was extended 250% in the PC-3 cohort and 300% in the H1299 cohort. Optimization of number and concentration of LNP doses should allow for long term control of both localized and systemic disease. In conclusion, we describe a novel LNP gene therapy approach for the treatment of cancer with high selectivity for tumors with dysregulated p53 transcriptional activation. This approach has the potential to provide a highly efficacious alternative to current therapies for localized and advanced solid tumors. Citation Format: Douglas Wilson Brown, Arun Raturi, Prakash Bhandari, Deborah Sosnowski, Liliya Grin, Ping Wee, Hector Vega, Jennifer Gyoba, Maryam Hejazi, Jailal Ablack, Matthew Scholz, John D. Lewis. Selective ablation of solid tumors using a p53-targeted FAST-LNP gene therapy [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 4069.

Quantitative Phosphoproteomics Reveals Cell Alignment and Mitochondrial Length Change under Cyclic Stretching in Lung Cells.

Lung cancer is a leading cause of death. Most previous studies have been based on traditional cell-culturing methods. However, lung cells are periodically subjected to mechanical forces during breathing. Understanding the mechanisms underlying the cyclic stretching induced in lung cells may be important for lung cancer therapy. Here, we applied cyclic stretching to stimulate the continual contraction that is present under physiological conditions in lung cells. We first uncovered the stretching-induced phosphoproteome in lung cancer cell line A549 and fibroblast cell line IMR-90. We identified 2048 and 2604 phosphosites corresponding to 837 and 1008 phosphoproteins in A549 and IMR-90, respectively. Furthermore, we combined our phosphoproteomics and public gene expression data to identify the biological functions in response to cyclic stretching. Interestingly, cytoskeletal and mitochondrial reorganization were enriched. We further used cell imaging analysis to validate the profiling results and found that this physical force changed cell alignment and mitochondrial length. This study not only reveals the molecular mechanism of cyclic stretching but also provides evidence that cell stretching causes cellular rearrangement and mitochondrial length change.

Clitoria ternatea L. petal bioactive compounds display antioxidant, antihemolytic and antihypertensive effects, inhibit α-amylase and α-glucosidase activities and reduce human LDL cholesterol and DNA induced oxidation.

The purpose of this study was to use a statistical approach to optimise the experimental conditions regarding the extraction of bioactive compounds, and to analyse the in vitro functional properties of crude lyophilized extracts (CLE) and partially purified (PPE) extracts of Clitoria ternatea petals. The results showed that the factors of temperature and time influenced the extraction of phenolic compounds, antioxidant activity and the physicochemical parameters. Simultaneous optimisation showed that the same levels of bioactive compounds were extracted when using temperatures from 11.7 to 68.3 °C and times from 8.47 to 51.12 min. Principal component analysis revealed the experimental conditions that provided the extraction producing the highest level of phenolic content (40 °C/30 min). The CLE showed antimicrobial activity; protective effect against hemolysis of erythrocytes; inhibition of α-amylase, α-glucosidase and angiotensin-I-converting (ACE-I) enzymes; and inhibition of lipid peroxidation. The CLE and PPE demonstrated oxygen radical absorption capacity; inhibition of DNA strand scission; inhibition of LDL cholesterol oxidation; intracellular antioxidant activity against reactive oxygen species (>100 μg/mL); and no cytotoxicity (IC50, GI50 and LC50 > 900 μg/mL) against A549, HCT8 and IMR90 cell lines.

Abstract 344: Proteomic Analysis Revealing Association of hiPSC-exosomal Proteins in Epigenetic Inheritance

Background: Exosomes, as a mediator of cell-to-cell communication has regulatory effects on the genetic and epigenetic processes in the recipient cells. Although the ability of human induced pluripotent stem cells (hiPSCs) derived exosomes to participate in tissue repair is being increasingly recognized, the information inside the hiPSC-exosomes especially protein content is not completely identified yet. Method: Exosomes derived from human fibroblast cell line IMR90 and IMR90 derived hiPSC cell line iPS (IMR90)-4 were collected using differential ultracentrifugation, then the exosomal proteins were identified by mass spectrometry analysis. The proteomics results were confirmed by western blot. To validate the role hiPSC-exosome in epigenetic processes, the H9C2 cells were treated with both types of exosomes and the histone epigenetic state were analyzed. Results: Gene ontology (GO) analysis of the exosome proteome showed that the hiPSC-exosome protein content had statistically significant enrichment in epigenetic modification such as acetylation, phosphorylation and methylation. The proteomic analysis indentified 6 epigenetic enzymes (UBE2N, DNMT3B, AURKB, CARM1, HDAC2 and HAT1) which only expressed in hiPSC-exosomes whereas 2 enzymes (PRMT5 and PRMT1) showed increased level in hiPSC-exosomes. The differential expressions of HDAC2 and PRMT5 were validated by western blot. Furthermore, H9C2 cell treated with hiPSC-exosomes showed a significant decrease in pan-acetylation level of H4 compared with IMR90-exosomes. Conclusion: In a global analysis presented here, hiPSC-exosomal proteins have been found to be associated with mammalian epigenetic inheritance, which provide important information for the exosome-based therapy.

PH-Responsive, Functionalizable Spyrocyclic Polycarbonate: A Versatile Platform for Biocompatible Nanoparticles.

Polymeric nanoparticles are widely investigated to enhance the selectivity of therapeutics to targeted sites, as well as to increase circulation lifetime and water solubility of poorly soluble drugs. In contrast to the encapsulation of the cargo into the nanostructures, the conjugation directly to the polymer backbone allows better control on the loading and selective triggered release. In this work we report a simple procedure to create biodegradable polycarbonate graft copolymer nanoparticles via a ring opening polymerization and subsequent postpolymerization modification strategies. The polymer, designed with both pH-responsive acetal linkages and a norbornene group, allows for highly efficient postpolymerization modifications through a range of chemistries to conjugate imaging agents and solubilizing arms to direct self-assembly. To demonstrate the potential of this approach, polycarbonate-based nanoparticles were tested for biocompatibility and theirability to be internalized in A549 and IMR-90 cell lines.

Posttranscriptional Regulation in Adenovirus Infected Cells.

A deeper understanding of how viruses reprogram their hosts for production of progeny is needed to combat infections. Most knowledge on the regulation of cellular gene expression during adenovirus infection is derived from mRNA studies. Here, we investigated the changes in protein expression during the late phase of adenovirus type 2 (Ad2) infection of the IMR-90 cell line by stable isotope labeling in cell culture with subsequent liquid chromatography-high resolution tandem mass spectrometric analysis. Two biological replicates of samples collected at 24 and 36 h post-infection (hpi) were investigated using swapped labeling. In total, 2648 and 2394 proteins were quantified at 24 and 36 hpi, respectively. Among them, 659 and 645 were deregulated >1.6-fold at the two time points. The protein expression was compared with RNA expression using cDNA sequencing data. The correlation was surprisingly low (r = 0.3), and several examples of posttranscriptional regulation were observed; e.g., proteins related to carbohydrate metabolism were up-regulated at the protein level but unchanged at the RNA level, whereas histone proteins were down-regulated at the protein level but up-regulated at the RNA level. The deregulation of cellular gene expression by adenovirus is mediated at multiple levels and more complex than hitherto believed.

Prior hypoxia prevents downregulation of ACE-2 by hyperoxia in fetal human lung fibroblasts

ABSTRACTPurpose/Aim of Study: The renin angiotensin system is involved in experimentally induced lung fibrosis. Angiotensin (ANG)-II is profibrotic. Angiotensin converting enzyme-2 (ACE-2) cleaves ANG-II and is thus protective. ACE-2 has recently been reported to be significantly decreased under hyperoxic conditions. Hyperoxia is linked to Bronchopulmonary Dysplasia and lung fibrosis. Fetal lung cells normally do not undergo fibrotic changes with physiologic hypoxemia. We hypothesized that hypoxia prior to hyperoxic exposure in fetal lung fibroblasts (IMR-90 cell line) might be protective by preventing ACE-2 downregulation. Materials and Methods: IMR-90 cells were exposed to hypoxia (1%O2/99%N2) followed by hyperoxia (95%O2/5%CO2) or normoxia (21%O2) in vitro. Cells and culture media were recovered separately for assays of ACE-2, TNF-α-converting enzyme (TACE), αSmooth muscle actin (αSMA)—myofibroblast marker-, N-cadherin, and β-catenin immunoreactive protein. Results: ACE-2 significantly increased when IMR-90 were hypoxic prior to hyperoxic exposure with no recovery. In contrast to hyperoxia alone, ACE-2 did not decrease when IMR-90 were hypoxic prior to hyperoxic exposure with recovery. TACE/ADAM17 protein and mRNA were significantly decreased under these conditions. αSMA N-cadherin, and β-catenin proteins were significantly decreased with or without normoxic recovery. Conclusions: Hypoxia prior to hyperoxic exposure of fetal lung fibroblasts prevented ACE-2 downregulation and decreased ADAM17/TACE protein and mRNA. αSMA, N-cadherin, and β-catenin were also significantly decreased under these conditions.

Detection of differentially methylated regions from whole-genome bisulfite sequencing data without replicates.

DNA methylation is an important epigenetic modification involved in many biological processes and diseases. Recent developments in whole genome bisulfite sequencing (WGBS) technology have enabled genome-wide measurements of DNA methylation at single base pair resolution. Many experiments have been conducted to compare DNA methylation profiles under different biological contexts, with the goal of identifying differentially methylated regions (DMRs). Due to the high cost of WGBS experiments, many studies are still conducted without biological replicates. Methods and tools available for analyzing such data are very limited.We develop a statistical method, DSS-single, for detecting DMRs from WGBS data without replicates. We characterize the count data using a rigorous model that accounts for the spatial correlation of methylation levels, sequence depth and biological variation. We demonstrate that using information from neighboring CG sites, biological variation can be estimated accurately even without replicates. DMR detection is then carried out via a Wald test procedure. Simulations demonstrate that DSS-single has greater sensitivity and accuracy than existing methods, and an analysis of H1 versus IMR90 cell lines suggests that it also yields the most biologically meaningful results. DSS-single is implemented in the Bioconductor package DSS.

PCTAIRE1/PCTK1/CDK16: a new oncotarget?

PCTAIRE1/PCTK1/CDK16: a new oncotarget?

Cell context dependent p53 genome-wide binding patterns and enrichment at repeats.

The p53 ability to elicit stress specific and cell type specific responses is well recognized, but how that specificity is established remains to be defined. Whether upon activation p53 binds to its genomic targets in a cell type and stress type dependent manner is still an open question. Here we show that the p53 binding to the human genome is selective and cell context-dependent. We mapped the genomic binding sites for the endogenous wild type p53 protein in the human cancer cell line HCT116 and compared them to those we previously determined in the normal cell line IMR90. We report distinct p53 genome-wide binding landscapes in two different cell lines, analyzed under the same treatment and experimental conditions, using the same ChIP-seq approach. This is evidence for cell context dependent p53 genomic binding. The observed differences affect the p53 binding sites distribution with respect to major genomic and epigenomic elements (promoter regions, CpG islands and repeats). We correlated the high-confidence p53 ChIP-seq peaks positions with the annotated human repeats (UCSC Human Genome Browser) and observed both common and cell line specific trends. In HCT116, the p53 binding was specifically enriched at LINE repeats, compared to IMR90 cells. The p53 genome-wide binding patterns in HCT116 and IMR90 likely reflect the different epigenetic landscapes in these two cell lines, resulting from cancer-associated changes (accumulated in HCT116) superimposed on tissue specific differences (HCT116 has epithelial, while IMR90 has mesenchymal origin). Our data support the model for p53 binding to the human genome in a highly selective manner, mobilizing distinct sets of genes, contributing to distinct pathways.

Prognostic significance of SOX18 expression in non-small cell lung cancer.

Recent studies have demonstrated the involvement of SOX18 transcription factor in blood and lymphatic vessel development, as well as in wound healing processes. SOX18 expression has been noted in cancer cells of various tumours, including lung cancer. However, the exact role of SOX18 expression in non-small cell lung cancer (NSCLC) remains to be determined. The present study, therefore, assessed its expression in 198 cases of NSCLC, consisting of 94 adenocarcinomas (AC), 89 squamous cell carcinomas (SQC) and 15 large cell carcinomas (LCC). The analysis utilized immunohistochemistry (IHC) and, in 42 cases, molecular methods. SOX18 expression was also determined in NSCLC cell lines (NCI-H1703, NCI-H522 and A549) and in normal lung fibroblasts (IMR-90). SOX18 was found to be expressed in nuclei, as well as in the cytoplasm of cancer cells, in the majority of studied cases. SOX18 mRNA expression was significantly lower in NSCLC than in non-malignant lung tissue (p<0.0001). However, SOX18 protein expression levels were higher in NSCLC tissues (p<0.005) and in the examined lung cancer cell lines. No SOX18 expression was noted in the IMR-90 cell line. In paraffin sections, a positive correlation between the Ki-67 antigen and nuclear SOX18 expression (r=0.17, p<0.05) was noted. In univariate survival analysis, cytoplasmic SOX18 expression correlated with poor patient outcome in the whole study and in AC cohorts (both p<0.05). Based on these results, SOX18 may be involved in the progression of NSCLC.

Subtelomeric CTCF and cohesin binding site organization using improved subtelomere assemblies and a novel annotation pipeline

Mapping genome-wide data to human subtelomeres has been problematic due to the incomplete assembly and challenges of low-copy repetitive DNA elements. Here, we provide updated human subtelomere sequence assemblies that were extended by filling telomere-adjacent gaps using clone-based resources. A bioinformatic pipeline incorporating multiread mapping for annotation of the updated assemblies using short-read data sets was developed and implemented. Annotation of subtelomeric sequence features as well as mapping of CTCF and cohesin binding sites using ChIP-seq data sets from multiple human cell types confirmed that CTCF and cohesin bind within 3 kb of the start of terminal repeat tracts at many, but not all, subtelomeres. CTCF and cohesin co-occupancy were also enriched near internal telomere-like sequence (ITS) islands and the nonterminal boundaries of subtelomere repeat elements (SREs) in transformed lymphoblastoid cell lines (LCLs) and human embryonic stem cell (ES) lines, but were not significantly enriched in the primary fibroblast IMR90 cell line. Subtelomeric CTCF and cohesin sites predicted by ChIP-seq using our bioinformatics pipeline (but not predicted when only uniquely mapping reads were considered) were consistently validated by ChIP-qPCR. The colocalized CTCF and cohesin sites in SRE regions are candidates for mediating long-range chromatin interactions in the transcript-rich SRE region. A public browser for the integrated display of short-read sequence–based annotations relative to key subtelomere features such as the start of each terminal repeat tract, SRE identity and organization, and subtelomeric gene models was established.

손상된 불가사리 추출물 흡입이 포름알데히드 노출에 의한 항염증 작용에 미치는 효과

포름알데히드는 주로 산업용으로 사용되는 독성이 강한 물질로서 생체에 노출 시 염증, 산화적 스트레스, 알레르기 반응, 나아가서 암을 유발하게 된다. 불가사리에서 추출되는 saponin은 sulphated sterol glycosides 등으로서 인삼 saponin (ginsenoside)들과 화학적 구조가 비슷하여 항균 및 세포독성을 비롯한 미생물을 괴사시키는 기능을 가지고 있음이 보고되었다. 또한 손상된 불가사리의 조직에는 염증성 사이토카인이 발현되는데 TGF-<TEX>${\beta}$</TEX>의 증가로 손상 받은 부위를 빠르게 치료, 재생하게 된다. 본 실험은 조직손상 후 8일 된 불가사리 추출물과 포름알데히드를 IMR-90에 처리했을 때와 포름알데히드에 노출시킨 ICR 마우스에 불가사리 추출물을 흡입시켰을 때 염증성 폐 손상의 방어 및 치료에 미치는 효과를 알아보았다. 포름알데히드에 노출된 IMR-90에 조직손상 후 8일이 경과한 불가사리 추출물을 처리한 결과, 세포의 유의한 증식을 보였으며 염증 조절에 영향을 주는 TNF-<TEX>${\alpha}$</TEX>, NF-<TEX>${\kappa}B$</TEX>의 발현억제 및 <TEX>$I{\kappa}-B{\alpha}$</TEX>의 합성을 촉진시켰다. 포름알데히드에 노출된 ICR 마우스에 조직손상 후 8일이 경과한 불가사리 추출물을 처리한 군에서 체중의 증가, 항산화 효소의 증가, 지질과산화 작용의 감소, surfactant protein A의 증가, TNF-<TEX>${\alpha}$</TEX> 및 NF-<TEX>${\kappa}B$</TEX>의 발현억제, <TEX>$I{\kappa}-B{\alpha}$</TEX> 발현의 증가와 조직재생에 우수한 효과를 보였다. 이러한 결과는 포름알데히드로 인한 세포 및 폐 손상을 불가사리 추출물의 항염증 작용을 통하여 억제하거나 완화시킴을 확인할 수 있다. Formaldehyde (FA) is widely used in industries, and it is an indoor and outdoor pollutant. Exposure to FA may cause inflammation and respiratory oxidative stress. Studies have demonstrated that FA can cause cancer in animal models. During the regeneration process of injured starfish (Asterina pectinifera), several changes have been observed in the expression of cytokines. In particular, higher TGF-<TEX>${\beta}1$</TEX> expression has been detected in arm cut starfish extract after eight days. The current study was designed to elucidate the in-vitro and the in-vivo pharmacological effects of starfish extract on FA exposure. We investigated the protective effects of intact starfish extract and arm cut starfish extract on an IMR-90 cell line and on mouse lung injury in response to FA exposure. In the presence of FA, inhalation of the arm cut starfish extract was associated with more promising cell proliferation, TNF-<TEX>${\alpha}$</TEX>, NF-<TEX>${\kappa}B$</TEX> decrement, and <TEX>$I{\kappa}-B{\alpha}$</TEX> increment. In the experimental group, the pulmonary structure of the arm cut starfish extract-treated group in the presence of FA exposure was similar to the control group, whereas the FA exposure group showed damage to the pulmonary structure. Moreover, the arm cut starfish extracts was more effective than the intact starfish extracts in terms of the expression of TNF-<TEX>${\alpha}$</TEX>, NF-<TEX>${\kappa}B$</TEX>, <TEX>$I{\kappa}-B{\alpha}$</TEX>, and surfactant protein A. The results obtained in this study demonstrate that arm cut starfish extracts are more effective in protecting pulmonary structure and function against FA exposure than intact starfish extracts.