Retinoic acid signaling pathway plays a role in regulating vertebrate development, cell differentiation, and homeostasis. As a key enzyme that catalyzes the oxidation of retinal to retinoic acid, aldehyde dehydrogenase 1 family member A2 (Aldh1a2) is involved in cardiac development, while whether it functions in heart diseases remains to be studied. In this study, we infected primary cardiomyocytes with adenovirus overexpressing Aldh1a2 (Ad-Aldh1a2) to explore the effects of Aldh1a2 overexpression on the biological function of cardiomyocytes. The results showed that the infection with Ad-Aldh1a2 realized the overexpression of Aldh1a2 in cardiomyocytes. Compared with the control group infected with Ad-GFP, the cardiomyocytes infected with Ad-Aldh1a2 showcased significantly increased size and up-regulated expression levels of the atrial natriuretic factor gene (ANF), brain natriuretic peptide gene (BNP), and β-myosin heavy chain (β-MHC). In addition, 5-ethynyl-2'-deoxyuridine (EdU) incorporation assay demonstrated that Aldh1a2 overexpression increased the proportion of cardiomyocytes with positive EdU signals and upregulated the expression levels of proliferation-related genes cyclin D2 (Ccnd2) and budding uninhibited by benzimidazole 1 (Bub1). The above data indicated that overexpression of Aldh1a2 induced hypertrophic growth and proliferation of cardiomyocytes. This study provides a basis for further understanding the function of Aldh1a2 in heart diseases and developing therapies for heart diseases.
Myocytes, Cardiac/cytology* ; Animals ; Cell Proliferation ; Aldehyde Dehydrogenase 1 Family/metabolism* ; Rats ; Retinal Dehydrogenase/metabolism* ; Adenoviridae/metabolism* ; Cells, Cultured ; Rats, Sprague-Dawley ; Cardiomegaly/metabolism* ; Up-Regulation ; Aldehyde Dehydrogenase, Mitochondrial

OBJECTIVETo investigate the expression of ALDH1, CXCR4 and E-cadherin in gastric carcinoma and their roles in lymphatic metastasis.

METHODSSurgical specimens from 127 cases of gastric carcinoma were examined for expressions of ALDH1, CXCR4 and E-cadherin immuohistochemistry with 60 adjacent tissues as control. The associations of ALDH1, CXCR4 and E-cadherin with the clinicopathological pfeatures, 5-year survival rate and lymph node metastasis of the patients were analyzed.

RESULTSALDH1, CXCR4 and E-cadherin were positive in 57.5% (73/127), 63.8% (81/127), and 36.2% (46/127) of the gastric carcinoma tissues, respectively, showing significant differences from the rates in the adjacent tissues (P<0.05). The expression of ALDH1 was significantly correlated with TNM stage and lymph node metastasis (P<0.05), CXCR4 was significantly correlated with the invasion depth, differentiation, TNM stage and lymph node metastasis of the tumor (P<0.05), and E-cadherin was significantly correlated with the invasion depth, differentiation and lymph node metastasis (P<0.05). The positivity rates of ALDH1, CXCR4 and E-cadherin were higher in cases with lymph node metastasis than in those without metastasis. E-cadherin expression was inversely correlated with ALDH1 and CXCR4 expression, and the latter two were positively correlated (P<0.001). Overexpressions of ALDH1 and CXCR4 and a decreased expression of E-cadherin were all related to a poor prognosis of the patients (P<0.05). The expressions ofALDH1, CXCR4 and E-cadherin were all independent prognostic factors of gastric carcinoma.

CONCLUSIONThe expressions of ALDH1, CXCR4 and E-cadherin are associated with the invasion, metastasis and prognosis of gastric carcinoma, and their combined detection provides important evidence for predicting the progression and prognosis of gastric carcinoma.

Cadherins ; genetics ; metabolism ; Carcinoma ; genetics ; metabolism ; Disease Progression ; Humans ; Isoenzymes ; genetics ; metabolism ; Lymphatic Metastasis ; Prognosis ; Receptors, CXCR4 ; genetics ; metabolism ; Retinal Dehydrogenase ; genetics ; metabolism ; Stomach Neoplasms ; genetics ; metabolism ; Survival Rate

AC133 Antigen ; Animals ; Antigens, CD ; metabolism ; Biomarkers, Tumor ; metabolism ; Breast Neoplasms ; metabolism ; pathology ; CD24 Antigen ; metabolism ; CD55 Antigens ; metabolism ; Female ; Gangliosides ; metabolism ; Glycoproteins ; metabolism ; Hedgehog Proteins ; metabolism ; Humans ; Hyaluronan Receptors ; metabolism ; Isoenzymes ; metabolism ; Neoplastic Stem Cells ; metabolism ; Octamer Transcription Factor-3 ; metabolism ; Peptides ; metabolism ; Receptors, Notch ; metabolism ; Retinal Dehydrogenase ; metabolism ; Signal Transduction ; Wnt Signaling Pathway

OBJECTIVETo investigate the expression pattern of CD133 and ALDH1 in colorectal cancer cells line Colo205 cultured in serum-free medium (SFM) containing recombinant human epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF).

METHODSColo205 cells were cultured in serum-free medium (SFM) containing human recombinant EGF and bFGF or in serum-supplemented medium (SSM). The expression of CD133 was analyzed in both groups, and CD133(+) and CD133(-) cells sorted from the SFM group using flow cytometry and observed microscopically for their growth status. The expression of CD133 and ALDH1 in CD133(+) cells and CD133(-) cells was detected by immunofluorescence assay. CD133(+) cells and CD133(-) cells were then injected subcutaneously into NOD/SCID mice and the expression of ALDH1 in the tumor tissues was detected by immunohistochemistry.

RESULTSThe cells in SFM group showed a significantly higher percentage of CD133(+) cells than those in SSM group (P<0.05). In SFM, CD133(+) cells were capable of forming tumor spheres while CD133(-) cells could not; CD133(+)cells strongly expressed CD133 and ALDH1 and CD133(-) cells did not. In mice, tumors generated by CD133(+) cells, but not by CD133(-) cells, positively expressed ALDH1.

CONCLUSIONSCD133(+) Colo205 colorectal cancer cells in SFM containing human recombinant EGF and bFGF can form tumor spheres and strongly express ALDH1. ALDH1 may be one of the candidate markers of colorectal cancer stem cells.

AC133 Antigen ; Animals ; Antigens, CD ; metabolism ; Cell Culture Techniques ; Cell Line, Tumor ; Colorectal Neoplasms ; metabolism ; Culture Media, Serum-Free ; Glycoproteins ; metabolism ; Humans ; Isoenzymes ; metabolism ; Mice ; Mice, Inbred NOD ; Mice, SCID ; Peptides ; metabolism ; Retinal Dehydrogenase ; metabolism

Abstract: The first-line drug artemisinin is widely used against malaria. Commercially available artemisinin is extracted from plants. However, the lack of sufficient raw material, artemisinin and the cost associated with the drug's manufacture have limited the supply of ACT to most malaria sufferers in the Developing World. As such, it is important to develop a low cost, fine to environment and high-quality method to supply sufficient and reliable quantities of artemisinin in the future. The field of synthetic biology, which utilizes cell factories to manipulate microbial metabolism to enhance the production of artemisinin and its intermediates, has a particularly strong impact by providing new platforms for chemical production. After a brief introduction of the artemisinin biosynthetic pathway, the present review focuses on the introduction of artemisinin biosynthetic genes, such as the genes encoding amorpha-4, 11-diene monooxygenase, NADPH: cytochrome P450 oxidoreductase, artemisinic aldehyde delta 11(13) reductase and aldehyde dehydrogenase. The review also addresses general considerations for potential contributions of synthetic biology to artemisinin production, with an emphasis on factors influencing interest compounds production in chassis cells.
Antimalarials ; metabolism ; supply & distribution ; Artemisinins ; metabolism ; supply & distribution ; Biosynthetic Pathways ; Cytochrome P-450 Enzyme System ; genetics ; Escherichia coli ; metabolism ; Gene Dosage ; Genetic Engineering ; Isoenzymes ; genetics ; RNA Nucleotidyltransferases ; genetics ; Retinal Dehydrogenase ; genetics ; Saccharomyces cerevisiae ; metabolism ; Synthetic Biology

OBJECTIVE: To determine the apoptosis-inducing effect of ultraviolet light (UV) on human lens epithelial cell (HLEC) and to explore the involvement of changes in ALDH1 folowing UV radiation. METHODS: HLEC was exposed to the same UV light source and was subsequently divided into 6 groups according to UV radiation time of 0 (control group), 5, 10, 15, and 30 min. Apoptosis was detected by AO/EB staining. Changes of ALDH1 in HLEC were detected by immunohistochemical staining and Western blot. RESULTS: The intensity of immunohistochemical staining and the rate of positive cells decreased with increase of UV time (P<0.05). The rate of positive ALDH1 cells was negatively correlated with the rate of apoptosis (r= -0.92, P<0.05). Western blot showed the integrated absorbance values significantly decreased with the increase of UV time (P<0.05). CONCLUSION ALDH1 in HLEC decreases with an increase of UV exposure, which may be related to UV induced apoptosis of HLEC.
Aldehyde Dehydrogenase 1 ; Apoptosis ; radiation effects ; Cells, Cultured ; Epithelial Cells ; cytology ; metabolism ; radiation effects ; Humans ; Isoenzymes ; genetics ; metabolism ; Lens, Crystalline ; cytology ; Retinal Dehydrogenase ; genetics ; metabolism ; Ultraviolet Rays ; adverse effects

OBJECTIVETo detect the expression of aldehyde dehydrogenase 1 in human laryngeal cancer cells in vitro, and to explore whether it can be used as a marker of stem cells in human laryngeal cancer.

METHODSFluorescence staining and flow cytometry were used to detect the expression of ALDH1 in a human laryngeal cancer Hep-2 cell line, and fluorescence activated cells sorting was used to separate ALDH1(br) cells. ALDH1 tumor cells were cultured and their ability of proliferation and differentiation was observed in vitro.

RESULTSThe expression of ALDH1 in Hep-2 cells was different. The number of cells highly expressing ALDH1 was 2.9% ± 0.6%. Compared with ALDH1(low) cells and unsorted cells, ALDH1(br) cells exhibited increased proliferation ability. In serum-containing RPM I1640 culture medium, the proportion of ALDH1(br) cells was decreasing as days passed. The percentage of ALDH1(br) cells decreased from 94.2% ± 3.8% to the level before sorting. The ALDH1(br) cells demonstrated enhanced tumorigenic ability in nude mice.

CONCLUSIONSIn the laryngeal cancer Hep-2 cell line, the highly ALDH1-expressing cells show a strong ability of differentiation, proliferation and tumorigenesis. It indicates that ALDH1 can be used as a new marker of stem cells of laryngeal cancer cells.

Animals ; Biomarkers, Tumor ; metabolism ; Cell Culture Techniques ; Cell Differentiation ; Cell Line, Tumor ; Cell Proliferation ; Cell Separation ; Flow Cytometry ; methods ; Humans ; Isoenzymes ; metabolism ; Laryngeal Neoplasms ; enzymology ; pathology ; Male ; Mice ; Mice, Nude ; Neoplasm Transplantation ; Neoplastic Stem Cells ; enzymology ; Random Allocation ; Retinal Dehydrogenase ; metabolism ; Tumor Burden

OBJECTIVETo isolate breast cancer stem cells from breast cancer patients and identify their biological characteristics.

METHODSMammospheric cells were purified and enriched from the tumor tissues of breast cancer patients using mammosphere culture. Their expressions of CD44 and CD24 were analyzed by flow cytometry, and ALDH1, ESA and Oct4 expressions were determined by Western Blotting. The primary mammospheric and adherent cells, at the density of 2×10(4), 2×10(5) or 2×10(6), were inoculated into NOD/SCID mice to observe their tumorigenic and metastatic activities.

RESULTSWith mammosphere culture method, 62.36% of the mammospheric cells showed CD44(+)/CD24(-/low) phenotype. The expressions of ALDH1, ESA and Oct4 in the mammospheric cells were significantly higher than those in the adherent culture-derived breast cancer cells (P<0.05). Primary mammospheric cells were at least 100-fold more tumorigenic than the adherent cells; the mammospheric cells were associated with liver or lung metastases, but the adherent cells were not.

CONCLUSIONMammosphere culture can be employed to obtain breast cancer stem cells from the tumor tissues of breast cancer patients.

Adult ; Animals ; Aryldialkylphosphatase ; metabolism ; Breast Neoplasms ; pathology ; CD24 Antigen ; metabolism ; Cell Culture Techniques ; methods ; Female ; Humans ; Hyaluronan Receptors ; metabolism ; Isoenzymes ; metabolism ; Mice ; Mice, Inbred NOD ; Mice, SCID ; Middle Aged ; Neoplastic Stem Cells ; cytology ; metabolism ; Octamer Transcription Factor-3 ; metabolism ; Primary Cell Culture ; Retinal Dehydrogenase ; metabolism

OBJECTIVE: To establish the 2-dimensional electrophoresis (2-DE) map in colonic mucosa in sub-healthy people with shapeless stool and healthy people, to identify the differential proteins by matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS), and to provide theoretical basis for the pathogenesis of intestinal mucosa in sub-healthy people with shapeless stool. METHODS: Two-DE was used to separate the total proteins from the intestinal mucosa in sub-healthy people (the sub-health group) with the shapeless stool and healthy volunteers (the control group). ImageMaster 2D Elite soft was applied to analyze the 2-DE images, and the differentially expressed protein spots between the 2 groups were identified by MALDI-TOF-MS, protein bank and information technique. RESULTS: We analyzed the average maps and obtained 517 protein spots in the sub-healthy group and 535 protein spots in the control group. Between the sub-healthy group and the control group, the mean of 366 protein spots was matched, and the matching rate was 70.79%. Ten differential protein spots were screened by MALDI-TOF-MS, and 8 were identified. Five out of the 8 spots were significantly decreased, while 3 out of the 8 were significantly increased. CONCLUSION The proteomic expression in colonic mucosa of people with shapeless stool is significantly different from that of healthy people. Eight differential proteins such as aldehyde dehydrogenase 1A1 isoform 1, 3-hydroxy-3-methylglutaryl-coenzyme A synthase 2 (mitochondrial), γ-actin, annexin A5 possibly involve in the pathogenesis of sub-healthy people with shapeless stool.
Actins ; metabolism ; Aldehyde Dehydrogenase ; metabolism ; Aldehyde Dehydrogenase 1 ; Annexin A5 ; metabolism ; Case-Control Studies ; Colon ; metabolism ; physiopathology ; Dyspepsia ; metabolism ; Electrophoresis, Gel, Two-Dimensional ; Female ; Humans ; Hydroxymethylglutaryl-CoA Synthase ; metabolism ; Intestinal Mucosa ; metabolism ; physiopathology ; Male ; Proteins ; genetics ; isolation & purification ; metabolism ; Proteome ; analysis ; Proteomics ; methods ; Retinal Dehydrogenase

Breast cancer stem cells are a group of undifferentiated cells with self-renewal and multidifferentiation potential. Chemotherapeutic and radiotherapeutic resistance, hypoxic resistance, high tumorigenicity, high cell invasion, and metastatic abilities are characteristics of these cells, which are responsible for breast cancer recurrence. Therefore, the correct sorting and identification of breast cancer stem cells is a primary step for research in this field. This article briefly describes the recent progress on sorting and identification technologies for breast cancer stem cells. Sorting technologies include the side population technique, technologies that depend on cell surface markers, ALDEFLUOR assays, and in situ detection. Identification technologies include mammosphere cultures, limited dilution in vitro, and in-vivo animal models. This review provides an important reference for breast cancer stem cell research, which will explore new methods for the treatment of patients with breast cancer.
AC133 Antigen ; ATP Binding Cassette Transporter, Sub-Family G, Member 2 ; ATP-Binding Cassette Transporters ; metabolism ; Aldehyde Dehydrogenase ; metabolism ; Animals ; Antigens, CD ; metabolism ; Breast Neoplasms ; pathology ; Female ; Flow Cytometry ; methods ; Glycoproteins ; metabolism ; Humans ; Hyaluronan Receptors ; metabolism ; Integrin alpha6 ; metabolism ; Integrin beta1 ; metabolism ; Integrin beta3 ; metabolism ; Isoenzymes ; metabolism ; Membrane Proteins ; metabolism ; Neoplasm Proteins ; metabolism ; Neoplastic Stem Cells ; metabolism ; pathology ; Peptides ; metabolism ; Retinal Dehydrogenase ; metabolism ; Side-Population Cells ; cytology ; metabolism