المؤلفون: Suling Liu, Qiaodan Deng, Lei Zhou, Dong Wang, Wei Ma, Lu Deng, Dandan Sheng

المصدر: Cell Biology and Toxicology

مصطلحات موضوعية: 0301 basic medicine, Gene isoform, Short Communication, Health, Toxicology and Mutagenesis, Population, Aldehyde dehydrogenase, Toxicology, Cell Line, Aldehyde dehydrogenase (ALDH), Immunoenzyme Techniques, 03 medical and health sciences, 0302 clinical medicine, Cancer stem cell, Neoplasms, ALDEFLUOR assay, medicine, Humans, BAAA, DEAB, education, education.field_of_study, biology, Genome, Human, Chemistry, Gene Expression Profiling, Cancer, Cell Biology, Aldehyde Dehydrogenase, medicine.disease, Enzyme assay, Isoenzymes, ALDH1A1, HEK293 Cells, 030104 developmental biology, Biochemistry, 030220 oncology & carcinogenesis, Neoplastic Stem Cells, Cancer stem cell (CSC), biology.protein, Stem cell

الوصف: Aldehyde dehydrogenases (ALDHs) defend intracellular homeostasis by catalyzing the conversion of toxic aldehydes into non-toxic carboxylic acids, which is of particular importance to the self-renewal of stem cells and cancer stem cells. The widely used ALDEFLUOR assay was initially designed to indicate the activity of ALDH1A1 in leukemia and has been demonstrated to detect the enzyme activity of several other ALDH isoforms in various cancer types in recent years. However, it is still elusive which isoforms, among the 19 ALDH isoforms in human genome, are the potential contributors in catalyzing ALDEFLUOR assay in different cancers. In the current study, we performed a screening via overexpressing each ALDH isoform to assess their ability of catalyzing ALDEFLUOR assay. Our results demonstrate that nine isoforms are active in ALDEFLUOR assay, whose overexpression significantly increases ALDH-positive (ALDH+) population. Further analysis of the expression of these active isoforms in various cancers reveals cancer-type specific expression patterns, suggesting that different cancer types may exhibit ALDEFLUOR activity through expression of specific active ALDH isoforms. This study strongly indicates that a detailed elucidation of the functions for each active ALDH isoform in CSCs is necessary and important for a profound understanding of the underlying mechanisms of ALDH-associated stemness. Electronic supplementary material The online version of this article (10.1007/s10565-018-9444-y) contains supplementary material, which is available to authorized users.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::ba6d92578447c48b2bf00e0b9f3c9c38Test
https://doi.org/10.1007/s10565-018-9444-yTest

المؤلفون: Houjie Liang, John W. Bullen, Daniele M. Gilkes, Hongxia Hu, Lisha Xiang, Gregg L. Semenza, Weibo Luo, Debangshu Samanta, Haiquan Lu, Naoharu Takano

المصدر: Oncotarget

مصطلحات موضوعية: medicine.medical_treatment, Breast Neoplasms, Mice, SCID, SIAH1, Protein Serine-Threonine Kinases, Transfection, Targeted therapy, Small hairpin RNA, Mice, 03 medical and health sciences, 0302 clinical medicine, Breast cancer, Aldefluor assay, Animals, Humans, Medicine, Neoplasm Metastasis, Triple-negative breast cancer, 030304 developmental biology, Cell Nucleus, 0303 health sciences, biology, business.industry, Tumor Suppressor Proteins, mammospheres, targeted therapy, medicine.disease, 3. Good health, Ubiquitin ligase, Phenotype, Oncology, 030220 oncology & carcinogenesis, Immunology, triple-negative breast cancer, MCF-7 Cells, Neoplastic Stem Cells, biology.protein, Cancer research, Heterografts, Female, Hypoxia-Inducible Factor 1, Signal transduction, Stem cell, basal-like breast cancer, business, Acyltransferases, Priority Research Paper, Signal Transduction, Transcription Factors

الوصف: // Lisha Xiang 1,2,3 , Daniele M. Gilkes 2,3 , Hongxia Hu 2,3 , Naoharu Takano 2,3,5 , Weibo Luo 2,3 , Haiquan Lu 2,3 , John W. Bullen 2,3 , Debangshu Samanta 2,3 , Houjie Liang 1 and Gregg L. Semenza 2,3,4 1 Department of Oncology and Southwest Cancer Center, Southwest Hospital, Third Military Medical University, Chongqing, China 2 Vascular Program, Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 3 McKusick-Nathans Institute of Genetic Medicine, Johns Hopkins University School of Medicine, Baltimore, MD 4 Departments of Pediatrics, Medicine, Oncology, Radiation Oncology, and Biological Chemistry, Johns Hopkins University School of Medicine, Baltimore, MD 5 Department of Biochemistry, School of Medicine, Keio University, Tokyo, Japan Correspondence: Gregg L. Semenza, email: // Keywords : Aldefluor assay, basal-like breast cancer, mammospheres, targeted therapy, triple-negative breast cancer Received : December 11, 2014 Accepted : December 12, 2014 Published : December 18, 2014 Abstract Intratumoral hypoxia, which is associated with breast cancer metastasis and patient mortality, increases the percentage of breast cancer stem cells (BCSCs) but the underlying molecular mechanisms have not been delineated. Here we report that hypoxia-inducible factor 1 (HIF-1) triggers the expression and activity of TAZ, a transcriptional co-activator that is required for BCSC maintenance, through two discrete mechanisms. First, HIF-1 binds directly to the WWTR1 gene and activates transcription of TAZ mRNA. Second, HIF-1 activates transcription of the SIAH1 gene, which encodes a ubiquitin protein ligase that is required for the hypoxia-induced ubiquitination and proteasome-dependent degradation of LATS2, a kinase that inhibits the nuclear localization of TAZ. Inhibition of HIF-1α, TAZ, or SIAH1 expression by short hairpin RNA blocked the enrichment of BCSCs in response to hypoxia. Human breast cancer database analysis revealed that increased expression (greater than the median) of both TAZ and HIF-1 target genes, but neither one alone, is associated with significantly increased patient mortality. Taken together, these results establish a molecular mechanism for induction of the BCSC phenotype in response to hypoxia.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::6ce54975077577724453f04d07096cdbTest
https://doi.org/10.18632/oncotarget.2997Test

المؤلفون: Ananth Annapragada, Nadarajah Vigneswaran, Varatharasa Thiviyanathan, Jean Wu, Qingshan Mu

المصدر: International Journal of Oncology

مصطلحات موضوعية: cancer stem cells, Cancer Research, Pathology, Chromosomal Proteins, Non-Histone, medicine.disease_cause, Metastasis, Mice, education.field_of_study, Nanog Homeobox Protein, Articles, Middle Aged, Cell cycle, 3. Good health, Gene Expression Regulation, Neoplastic, Isoenzymes, Oncology, Head and Neck Neoplasms, Carcinoma, Squamous Cell, Neoplastic Stem Cells, Female, Homeobox protein NANOG, medicine.medical_specialty, tumorspheres, Population, Biology, head and neck squamous cell carcinoma, Aldehyde Dehydrogenase 1 Family, stomatognathic system, Cancer stem cell, otorhinolaryngologic diseases, medicine, Animals, Humans, Aldefluor assay, xenograft, education, neoplasms, Homeodomain Proteins, Squamous Cell Carcinoma of Head and Neck, Proteins, Retinal Dehydrogenase, Cancer, medicine.disease, Xenograft Model Antitumor Assays, Head and neck squamous-cell carcinoma, stomatognathic diseases, Fanconi Anemia, Neoplasm Recurrence, Local, Carcinogenesis, Octamer Transcription Factor-3

الوصف: Fanconi anemia (FA) patients have an increased risk of head and neck squamous cell carcinoma (HNSCC) at a higher rate with no apparent risk factors. HNSCC of FA patients is an aggressive tumor characterized by multifocal origin, early metastases and frequent recurrences. Given that cancer stem cells (CSC) drive tumorigenesis, tumor recurrence and metastasis, in this study, we characterized the CSC population in FA and sporadic HNSCC. The Aldefluor assay was used to characterize and isolate CSC with high aldehyde dehydrogenase (ALDH) activity (ALDHpos) in cell lines derived from FA and sporadic HNSCC. Isolated ALDHpos and ALDHneg cells were examined for the expression of stemness genes using reverse transcription-polymerase chain reaction (RT-PCR) array. Tumor cell-derived FA and sporadic HNSCC were examined for their ability to form tumorspheres in vitro. Stem-like cell population in FA and sporadic HNSCC in human and mouse xenograft tumors were evaluated using ALDH isoform 1 (ALDH1) immunohistochemistry. FA-HNSCC cell lines harbor a greater proportion of ALDHpos cells (15–31%) compared to sporadic HNSCC (10%). Expression of Nanog, Oct-3/4 and Stella, molecular markers of undifferentiated embryonic stem (ES) cells were detected in the ALDHpos FA-HNSCC cells and not in the ALDHneg cells. FA-HNSCC cell lines revealed enhanced in vitro tumorsphere formation compared to sporadic HNSCC cells. A higher percentage of ALDH1pos tumor cells are noted in the human and mouse xenograft tumors of FA-HNSCC compared to sporadic HNSCC tumors. FA-HNSCC are highly enriched for CSC and may serve as a model to develop CSC-targeted therapies for HNSCC.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::2ec437e7535c4dc0a65372ff15859dccTest
https://doi.org/10.3892/ijo.2014.2677Test

المؤلفون: Jian Geng, Chin-Lee Wu, Xiao Li, Liyan Wan, Xiaoyan Bai

المصدر: Journal of Thoracic Oncology. 7:1235-1245

مصطلحات موضوعية: Male, Lung Neoplasms, Fluorescent Antibody Technique, Apoptosis, medicine.disease_cause, Tissue microarray, Metastasis, Immunoenzyme Techniques, Mice, Cell Movement, Carcinoma, Non-Small-Cell Lung, Lung, Cells, Cultured, Mice, Inbred BALB C, Reverse Transcriptase Polymerase Chain Reaction, Cell Cycle, Transfection, respiratory system, Flow Cytometry, Prognosis, medicine.anatomical_structure, Oncology, Lymphatic Metastasis, Carcinoma, Squamous Cell, Disease Progression, Female, Pulmonary and Respiratory Medicine, Blotting, Western, Mice, Nude, Adenocarcinoma, Biology, Real-Time Polymerase Chain Reaction, Aldehyde Dehydrogenase 1 Family, Cancer stem cell, ALDEFLUOR assay, Cell Adhesion, medicine, Carcinoma, Animals, Humans, RNA, Messenger, Lung cancer, ALDH1A1, Cell Proliferation, Neoplasm Staging, Retinal Dehydrogenase, Aldehyde Dehydrogenase, medicine.disease, Molecular biology, respiratory tract diseases, biology.protein, Carcinogenesis, Small interfering RNA transfection, Follow-Up Studies

الوصف: Introduction:Lung cancer contains a small population of cancer stem cells that contribute to its initiation and progression. We investigated the biological function and clinical significance of aldehyde dehydrogenase 1A1 (ALDH1A1) in non–small-cell lung carcinoma (NSCLC).Methods:ALDH1A1 assay or small interfering RNA transfection was employed to isolate ALDH1A1+ cells or knock down ALDH1A1 expression in H2087 cells, respectively. Biological functions of ALDH1A1+ and ALDH1A1 silenced cells were investigated using in vitro and in vivo methods. ALDH1A1 expression was analyzed using immunohistochemistry on tissue microarrays with 179 lung cancer tissues and 26 normal lung tissues.Results:The abilities of clone formation, proliferation, cell growth, and migration were increased in ALDH1A1+ and ALDH1A1 silenced cells. ALDH1A1+ lung cancer cells initiated tumors that resembled the histopathologic characteristics and heterogeneity of the parental lung cancer cells in mice. The silencing of ALDH1A1 expression in H2087 lung cancer cells inhibited cell proliferation and migration significantly. ALDH1A1 was expressed in 42% of normal lung tissues (11 of 26), with strong expression in the basal cells and globular cells of the normal bronchus and weak expression in the alveolar epithelial cells. Compared with normal lung tissues, 45% of NSCLC samples (81 of 179) were read as positive for ALDH1A1. Positive ALDH1A1 expression was correlated with patients’ smoking status (p = 0.022), lymph-node metastasis (p = 0.006), clinical stage (p = 0.004), and a decreased overall survival time (p < 0.001). Positive ALDH1A1 expression in lung cancer tissues was an independent prognostic factor for NSCLC (odds ratio = 5.232, p < 0.001).Conclusion:Elucidating the biological functions of ALDH1A1 could be helpful in studying lung tumorigenesis and for developing new therapeutic approaches.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::0196c7999195e7c7d6566c1c8cda81b6Test
https://doi.org/10.1097/jto.0b013e318257cc6dTest

المؤلفون: Kokorsch, Philipp

المساهمون: Lengerke, Claudia (Prof. Dr.)

مصطلحات موضوعية: CDCP1, Brustkrebs , Eierstockkrebs, ALDH, Aldefluor Assay

الوصف: In dieser Arbeit wurde die Bedeutung der ALDH Aktivität und CDCP1 Expression für Tumorzelllinien des Mammakarzinoms, des Lungenadenokarzinoms, und des Ovarialkarzinoms sowie für primäres ovariales Tumorgewebe untersucht. Beiden Markern ist eine Assoziation mit erhöhtem Metastasierungspotential sowie mit invasivem Wachstum und Resistenz gegenüber Chemotherapie gemein. Ziel war herauszufinden ob jeweils eine Assoziation zu Tumorstammzellpotential besteht. Eine Tumorstammzellassoziation ergab sich für beide Marker bei Leukämie und für die ALDH auch bei weiteren soliden humanen Tumoren, wie dem Mammakarzinom. Jedoch ist die funktionelle Rolle der ALDH nicht für jedes Gewebe gleichbedeutend oder sie ist noch unklar; aktuelle Daten zum Ovarialkarzinom sind widersprüchlich. Zunächst wurden sämtliche Zelllinien im Aldefluor-Assay, bzw. durch Verwendung des primären Antikörpers CUB2 auf ihre Markerexpression hin analysiert. Anschließ-end wurde das Tumorstammzellpotential der positiv markierten Zellpopulationen im Sphärenassay untersucht. Beim Ovarialkarzinom wurden zusätzlich Zellzyklusanaly-sen und Untersuchungen der Differenzierungskapazität durchgeführt. Es ergab sich, dass CDCP1 sowohl in der Durchflusszytometrie als auch im Sphä-renassay nicht geeignet ist umschriebene Populationen zu isolieren. Dagegen zeigte sich beim Mammakarzinom bei der ALDH positiven Population der Zelllinie SKBR-3 ein stärkeres Potential gegenüber der negativen Population Sphären zu bilden. Es konnte gezeigt werden, dass auch beim Ovarialkarzinom Sphärenbildung möglich ist und dass die Fähigkeit der Ovarosphärenbildung in Zusammenhang mit der ALDH Aktivität steht. Auch das Proliferationspotential der ovarialen ALDH positiven Popula-tion, im Fall von der Zelllinie OVCAR-3, unterscheidet sich signifikant von dem der ALDH negativen Population. Allerdings zeigte sich hier auch bei der ALDH negativen Population eine Differenzierung in Richtung ALDH positiver Zellen. Die Daten dieser Arbeit zeigen, dass CDCP1 nicht als Tumorstammzelmarker benut-zt werden kann. Anders verhält es sich bei der ALDH, deren erhöhte Aktivität, bei verschiedenen humanen Tumoren, wie dem Mammakarzinom mit Stammzellpotential assoziiert ist, was durch diese Arbeit belegt wurde. Der Sphären-Assay konnte für die ALDH positive Tumorzellpopulation des Ovarialkarzioms etabliert werden und dieser Population somit Stammzellpotential zugesprochen werden. Untersuchungen der Proliferationskapazität widersprechen dem Tumorstammzellcharakter der ALDH positiven Population beim Ovarialkarzinom jedoch. Definitiv kennzeichnet dieser funktionelle Marker Zellen mit erhöhter proliferativer Kapazität.

وصف الملف: application/pdf

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::a70e305dcb498c3f7e70d3ea0997e774Test
https://hdl.handle.net/10900/65995Test