1.Inhibitor of DNA-binding 4 contributes to the maintenance and expansion of cancer stem cells in 4T1 mouse mammary cancer cell line.
Se Jin PARK ; Ran Ju KIM ; Jeong Seok NAM
Laboratory Animal Research 2011;27(4):333-338
The cancer stem cell (CSC) hypothesis proposes that CSCs are the root of cancer. CSC-targeted therapies may prevent cancer relapse and provide more effective treatment. The expression of aldehyde dehydrogenase 1, as assessed by the Aldefluor assay, has been recognized as a marker of CSCs in breast cancer. Inhibitors of DNA-binding proteins (IDs) have an important role in stem cell differentiation. In this study, we examined IDs necessary for the regulation of stem properties in Aldefluorpos 4T1 cells. When the expression profile of IDs in Aldefluorneg and Aldefluorpos 4T1 cells was compared, qRT-PCR analysis showed that ID4 expression was highly upregulated in Aldefluorpos 4T1 cells. In addition, knockdown of ID4 expression suppressed the properties of CSCs, including their sphere-forming ability and side population phenotype. The findings suggest that ID4 may be a therapeutic target for the treatment of advanced breast cancer.
Aldehyde Dehydrogenase
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Animals
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Breast Neoplasms
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Cell Line
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DNA-Binding Proteins
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Isoenzymes
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Mice
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Neoplastic Stem Cells
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Phenotype
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Recurrence
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Retinal Dehydrogenase
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Stem Cells
2.NADPH Diaphorase Staining Retinal Cells in Streptozotocin-induced Diabetic Rat Retina.
Journal of the Korean Ophthalmological Society 1998;39(10):2377-2384
Nitric oxide(NO) is a free radical which serves a wide variety of functions on vascular tone, neurotransmission, immune cytotoxicity, and many others. Nitric oxide synthase(NOS) is the biosynthetic enzyme of NO and colocalized with NADPH diaphorase(NADPH-d) activity in many tissues. The author aimed to assess the changes that occur in this populations of neurons in the streptozotocin-induced diabetic rat where the retinal vasculature is known to be dysfunctional. The 8 rats was a diabetic group and the other 8 was a control group. Diabetes was induced with a single intraperitoneal injection of streptozotocin(65mg/kg). Four weeks later, the retina was flat mounted and stained with NADPH-d. Counting of the stained cells was made. There was a 20.6% decrease in the total number of positively staining cells in the retinas of the diabetic group(2532+/-192) compared with those of the control group(3188+/-176)(p<0.001). It is worth to suggest the close correlation between NO released from retinal neurons and the microcirculatory dysfunction in diabetic retinopathy.
Animals
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Diabetic Retinopathy
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Injections, Intraperitoneal
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NADP*
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NADPH Dehydrogenase*
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Neurons
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Nitric Oxide
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Rats*
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Retina*
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Retinal Neurons
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Retinaldehyde*
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Synaptic Transmission
3.OCT4 Expression Enhances Features of Cancer Stem Cells in a Mouse Model of Breast Cancer.
Laboratory Animal Research 2011;27(2):147-152
The cancer stem cell (CSC) hypothesis proposes that CSCs are responsible for metastasis and disease recurrence. Therefore, targeting CSCs has the potential to significantly improve outcomes for cancer patients. The OCT4 transcription factor gene is a master gene that plays a key role in the self-renewal and pluripotency of stem cells. In this study, we introduced an OCT4 reporting vector into 4T1 mouse breast cancer cells and sorted OCT4 high and OCT4 low cell populations. We then determined whether OCT4 expression is associated with maintenance and expansion of CSCs. We found that OCT4high 4T1 cells have an increased ability to form tumorsphere and a high expression of stem cell markers such as Sca-1, CD133, CD34, and ALDH1, when compared with OCT4low 4T1 cells. In addition, OCT4high 4T1 cells have greater tumorigenic potential in vivo. These findings suggest that OCT4 expression may be a useful target for stem cell-specific cancer therapy.
Animals
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Breast
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Breast Neoplasms
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Humans
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Isoenzymes
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Mice
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Neoplasm Metastasis
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Neoplastic Stem Cells
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Recurrence
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Retinal Dehydrogenase
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Stem Cells
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Transcription Factors
4.Advancement of studies on ALDH1 as a universal marker of stem cells.
Journal of Biomedical Engineering 2010;27(5):1183-1186
Aldehyde dehydrogenase 1 (ALDH1, ALDH1A1 or RALDH1), an enzyme responsible for the oxidation of intracellular aldehydes, was shown to have a function in the early differentiation of stem cells. Its activity shows promising potential as a universal marker for the identification and isolation of normal stem cells and cancer stem cells from multiple sources in a variety of tissue types. Herein, we review the available data reporting the utilization of ALDH1 activity as a means to identify and isolate normal stem cells and cancer stem cells (CSCs), and the potential diagnostic and therapeutic implications, with a special focus on the mammary gland and breast cancer. The research opportunity in this area of interest is emphasized.
Animals
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Biomarkers
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metabolism
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Biomarkers, Tumor
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metabolism
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Humans
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Isoenzymes
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metabolism
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Neoplastic Stem Cells
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metabolism
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Retinal Dehydrogenase
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metabolism
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Stem Cells
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metabolism
5.Effect of ultraviolet radiation on ALDH1 expression in human lens epithelial cells.
Jingming SHI ; Songbai JIA ; Xuan CHEN ; Luosheng TANG
Journal of Central South University(Medical Sciences) 2012;37(6):572-577
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
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Apoptosis
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radiation effects
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Cells, Cultured
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Epithelial Cells
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cytology
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metabolism
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radiation effects
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Humans
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Isoenzymes
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genetics
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metabolism
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Lens, Crystalline
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cytology
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Retinal Dehydrogenase
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genetics
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metabolism
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Ultraviolet Rays
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adverse effects
6.Low Retinal Dehydrogenase 1 (RALDH1) Level in Prepubertal Boys with Autism Spectrum Disorder: A Possible Link to Dopamine Dysfunction?.
Denis PAVĂL ; Florina RAD ; Răzvan RUSU ; Alexandru Stefan NICULAE ; Horaţiu Alexandru COLOSI ; Iuliana DOBRESCU ; Eleonora DRONCA
Clinical Psychopharmacology and Neuroscience 2017;15(3):229-236
OBJECTIVE: Retinal dehydrogenase 1 (RALDH1) is a cytosolic enzyme which acts both as a source of retinoic acid (RA) and as a detoxification enzyme. RALDH1 has key functions in the midbrain dopaminergic system, which influences motivation, cognition, and social behavior. Since dopamine has been increasingly linked to autism spectrum disorder (ASD), we asked whether RALDH1 could contribute to the autistic phenotype. Therefore, we investigated for the first time the levels of RALDH1 in autistic patients. To further assess the detoxification function of RALDH1, we also explored 4-hydroxynonenal protein adducts (4-HNE PAs) and reduced glutathione (GSH) levels. Moreover, considering the effect of testosterone on RALDH1 expression, we measured the second to fourth digit ratio (2D:4D ratio) for both hands, which reflects exposure to prenatal testosterone. METHODS: Male patients with ASD (n=18; age, 62.9±4.3 months) and healthy controls (n=13; age, 78.1±4.9 months) were examined. Erythrocyte RALDH1, serum 4-HNE PAs and erythrocyte GSH levels were measured using colorimetric assays, and digit lengths were measured using digital calipers. RESULTS: We found significantly lower (−42.9%) RALDH1 levels in autistic patients as compared to controls (p=0.032). However, there was no difference in 4-HNE PAs levels (p=0.368), GSH levels (p=0.586), or 2D:4D ratios (p=0.246 in the left hand, p=0.584 in the right hand) between healthy controls and autistic subjects. CONCLUSION: We concluded that a subset of autistic patients had a low RALDH1 level. These results suggest that low RALDH1 levels could contribute to the autistic phenotype by reflecting a dopaminergic dysfunction.
Autism Spectrum Disorder*
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Autistic Disorder*
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Cognition
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Cytosol
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Dopamine*
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Erythrocytes
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Glutathione
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Hand
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Humans
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Male
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Mesencephalon
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Motivation
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Phenotype
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Retinal Dehydrogenase*
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Retinaldehyde*
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Social Behavior
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Testosterone
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Tretinoin
7.Effects of 5-Aza-2'-deoxycitydine and trichostatin A on expression and apoptosis of ALDH1a2 gene in human bladder cancer cell lines.
Zheng-hua JU ; Min-gang YING ; Xing AI ; Tao-ping SHI ; Bao-jun WANG ; Chao WANG ; Guo-xi ZHANG ; Xu ZHANG
Chinese Journal of Surgery 2010;48(5):378-382
OBJECTIVETo study the effect on promoter de-methylation, expression of ALDH1a2 gene and cell apoptosis by treated with 5-Aza-dC and TSA in five human bladder cancer cell lines.
METHODSHuman bladder cancer cell lines RT-4, 253J, 5637, BIU-87 and T24 were cultured and treated with 5-Aza-dC and(or) TSA. The expression of the ALDH1a2 gene was detected by RT-PCR and Western blot. The methylation status of gene promoter was determined by MSP, and the cell cycle profile was established by flow cytometry.
RESULTSALDH1a2 was silenced in five human bladder cancer cell lines. Re-expression of ALDH1a2 was detected after treated with 5-Aza-dC alone or TSA in combination. ALDH1a2 transcript was marked in each cell lines combined with 5-Aza-dC and TSA treatment which showed a synergistic effect on expression of ALDH1a2 transcript. Early apoptotic was the main mode of apoptosis and death of human bladder cancer cell lines induced by 5-Aza-dC and TSA. The percentage of early apoptotic cells was 1.4% in control group and 2.8% in TSA group, however, 20.2% in 5-Aza-dC group and 33.8% in 5-Aza-dC + TSA group, respectively. The groups of TSA, 5-Aza-dC and 5-Aza-dC + TSA were significantly different from control group (P < 0.05).
CONCLUSIONSAberrant methylation of ALDH1a2 gene is the main cause for gene transcriptional inactivation. Re-expression of ALDH1a2 gene and cell apoptosis are detected after either treatment with 5-Aza-dC alone or in combination with TSA.
Apoptosis ; drug effects ; Azacitidine ; pharmacology ; Cell Line, Tumor ; Gene Expression Regulation, Neoplastic ; Humans ; Hydroxamic Acids ; pharmacology ; Retinal Dehydrogenase ; metabolism ; Urinary Bladder Neoplasms ; metabolism ; pathology
8.Proteomics of the colonic mucosa in sub-healthy people with shapeless stool.
Baiyun ZHONG ; Hui DENG ; Jingzhong LIAO ; Yunli ZHANG ; Weimin XIE ; Weidong LIU ; Sisi FENG
Journal of Central South University(Medical Sciences) 2011;36(9):817-822
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
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metabolism
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Aldehyde Dehydrogenase
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metabolism
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Aldehyde Dehydrogenase 1
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Annexin A5
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metabolism
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Case-Control Studies
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Colon
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metabolism
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physiopathology
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Dyspepsia
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metabolism
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Electrophoresis, Gel, Two-Dimensional
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Female
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Humans
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Hydroxymethylglutaryl-CoA Synthase
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metabolism
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Intestinal Mucosa
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metabolism
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physiopathology
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Male
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Proteins
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genetics
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isolation & purification
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metabolism
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Proteome
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analysis
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Proteomics
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methods
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Retinal Dehydrogenase
9.Expression of ALDH1, CXCR4 and E-cadherin in castric carcinoma and their roles in lymphatic metastasis.
Yan ZHAO ; Xin JIN ; Nan LI ; Jing LI ; Jun QIAN
Journal of Southern Medical University 2016;36(10):1390-1395
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
10.Research progresses in synthetic biology of artemisinin.
Jian-Qiang KONG ; Wei WANG ; Ke-Di CHENG ; Ping ZHU
Acta Pharmaceutica Sinica 2013;48(2):193-205
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
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metabolism
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supply & distribution
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Artemisinins
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metabolism
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supply & distribution
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Biosynthetic Pathways
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Cytochrome P-450 Enzyme System
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genetics
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Escherichia coli
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metabolism
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Gene Dosage
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Genetic Engineering
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Isoenzymes
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genetics
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RNA Nucleotidyltransferases
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genetics
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Retinal Dehydrogenase
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genetics
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Saccharomyces cerevisiae
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metabolism
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Synthetic Biology