1.Analysis of lysosomal membrane proteins exposed to melanin in HeLa cells.
Seung Hyuck BANG ; Dong Jun PARK ; Yang Hoon KIM ; Jiho MIN
Environmental Health and Toxicology 2016;31(1):e2016009-
OBJECTIVES: There have been developed to use targeting ability for antimicrobial, anticancerous, gene therapy and cosmetics through analysis of various membrane proteins isolated from cell organelles. METHODS: It was examined about the lysosomal membrane protein extracted from lysosome isolated from HeLa cell treated by 100 ppm melanin for 24 hours in order to find associated with targeting ability to melanin using by 2-dimensional electrophoresis. RESULTS: The result showed 14 up-regulated (1.5-fold) and 13 down-regulated (2.0-fold) spots in relation to melanin exposure. CONCLUSIONS: It has been found that lysosomal membrane proteins are associated with melanin to decolorize and quantity through cellular activation of lysosome.
Electrophoresis
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Genetic Therapy
;
HeLa Cells*
;
Humans
;
Lysosome-Associated Membrane Glycoproteins*
;
Lysosomes
;
Melanins*
;
Membrane Proteins
;
Organelles
2.Expression of Lysosomal Membrane Proteins LAMP1, TPC1 and TPC2 in Acute Myeloid Leukemia Cells and Its Clinical Significance.
Xiang WANG ; Bing-Qian LING ; Mei-Fang DAI ; Xing-Bing WANG
Journal of Experimental Hematology 2019;27(4):1046-1052
OBJECTIVE:
To investigate the relationship between the expression of lysosomal membrane proteins LAMP1, TPC1 and TPC2 in acute myeloid leukemia (AML) cells and clinical indications of AML and to explore the possible role in the genesis and development of AML and clinical significance.
METHODS:
Real-time quantitative PCR was used to detect the mRNA expression of LAMP1, TPC1 and TPC2 in AML cell lines (HL-60, NB4) and 57 patients with acute myeloid leukemia (including 44 initially treated patients and 13 relapsed and refractory patients). The relationship of mRNA expression levels with clinical indicators and post-chemotherapy remission was analyzed.
RESULTS:
Compared with CD34 hematopoietic stem cells (HSC), the expression levels of LAMP1 and TPC1 in AML cell lines HL-60 and NB4 significantly increased, while the expression level of TPC2 was not significantly different. The expression levels of LAMP1, TPC1 and TPC2 in bone marrow mononuclear cells (BMMNC) of AML patients were higher than those in normal human BMMNC (P<0.05), and the expression levels of LAMP1, TPC1 and TPC2 in CD34 primary AML cells(CD34 primary cells in the patient's bone marrow >90%) were also high. There was no significant difference in the expression of LAMP1, TPC1 and TPC2 between CD34HSC of patients with AML and relapsed/refractory patients (P>0.05). No correlation was found between age, sex and genotype and expression of membrane proteins (P>0.05). The expression levels of LAMP1 and TPC1 positively correlated with the number of white blood cells in peripheral blood of patients (P<0.01). LAMP1 and TPC2 were found to be associated with remission after a course of chemotherapy in newly diagnosed patients. Initially treated patients with high expression of LAMP1 in the bone marrow not easily relieved after one course of chemotherapy. Patients with high expression of TPC2 in the bone marrow more likely to be relieved after one course of chemotherapy.
CONCLUSION
The mRNA of the three membrane proteins are highly expressed in AML patients, and LAMP1 and TPC1 are risk factors for AML disease progression. High expression of TPC2 is beneficial for chemotherapy of patients with newly diagnosed AML.
Bone Marrow
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Bone Marrow Cells
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Hematopoietic Stem Cells
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Humans
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Leukemia, Myeloid, Acute
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Lysosome-Associated Membrane Glycoproteins
3.Lysosomal membrane protein Sidt2 knockout induces apoptosis of human hepatocytes in vitro independent of the autophagy-lysosomal pathway.
Jiating XU ; Mengya GENG ; Haijun LIU ; Wenjun PEI ; Jing GU ; Mengxiang QI ; Yao ZHANG ; Kun LÜ ; Yingying SONG ; Miaomiao LIU ; Xin HU ; Cui YU ; Chunling HE ; Lizhuo WANG ; Jialin GAO
Journal of Southern Medical University 2023;43(4):637-643
OBJECTIVE:
To explore the regulatory mechanism of human hepatocyte apoptosis induced by lysosomal membrane protein Sidt2 knockout.
METHODS:
The Sidt2 knockout (Sidt2-/-) cell model was constructed in human hepatocyte HL7702 cells using Crispr-Cas9 technology.The protein levels of Sidt2 and key autophagy proteins LC3-II/I and P62 in the cell model were detected using Western blotting, and the formation of autophagosomes was observed with MDC staining.EdU incorporation assay and flow cytometry were performed to observe the effect of Sidt2 knockout on cell proliferation and apoptosis.The effect of chloroquine at the saturating concentration on autophagic flux, proliferation and apoptosis of Sidt2 knockout cells were observed.
RESULTS:
Sidt2-/- HL7702 cells were successfully constructed.Sidt2 knockout significantly inhibited the proliferation and increased apoptosis of the cells, causing also increased protein expressions of LC3-II/I and P62(P < 0.05) and increased number of autophagosomes.Autophagy of the cells reached a saturated state following treatment with 50 μmol/L chloroquine, and at this concentration, chloroquine significantly increased the expressions of LC3B and P62 in Sidt2-/- HL7702 cells.
CONCLUSION
Sidt2 gene knockout causes dysregulation of the autophagy pathway and induces apoptosis of HL7702 cells, and the latter effect is not mediated by inhibiting the autophagy-lysosomal pathway.
Humans
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Lysosome-Associated Membrane Glycoproteins/metabolism*
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Autophagy
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Apoptosis
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Hepatocytes
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Lysosomes/metabolism*
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Chloroquine/pharmacology*
;
Nucleotide Transport Proteins/metabolism*
5.Advances in research of SCARB2 functions and related disorders.
Miao HE ; Zhen LIU ; Beisha TANG ; Junling WANG
Chinese Journal of Medical Genetics 2015;32(5):723-727
SCARB2 (scavenger receptor class B, member 2) is a lysosomal membrane glucoprotein, which is encoded by SCARB2 gene. It takes vital parts in the physiological and pathological processes including the transportation of beta-glucocerebrosidase to the lysosome, infection of EV71 and load-induced cardiac myocyte hypertrophy. This article has reviewed the molecular structure and functions of SCARB2 gene and its protein, as well as their relationship with diseases.
Hand, Foot and Mouth Disease
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genetics
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Humans
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Lysosome-Associated Membrane Glycoproteins
;
chemistry
;
genetics
;
physiology
;
Myoclonic Epilepsies, Progressive
;
genetics
;
Parkinson Disease
;
genetics
;
Receptors, Scavenger
;
chemistry
;
genetics
;
physiology
6.Expression of EV71-VP1, PSGL-1 and SCARB2 in Tissues of Infants with Brain Stem Encephalitis.
Ming LI ; Xiao-ping KONG ; Hong LIU ; Ling-xi CHENG ; Jing-lu HUANG ; Li QUAN ; Fang-yu WU ; Bo HAO ; Chao LIU ; Bin LUO
Journal of Forensic Medicine 2015;31(2):97-104
OBJECTIVE:
To understand the correlation of enterovirus 71 (EV71), P-selectin glycoprotein ligand-1 (PSGL-1), and scavenger receptor B2 (SCARB2) and to explore the possible pathway and mechanism of EV71 infection by observing the expression of EV71, PSGL-1 and SCARB2 in tissues of infants with brain stem encephalitis.
METHODS:
The organs and tissues of infants with EV71-VP1 positivity in their brain stems were chosen. Expression and distribution of EV71-VP1, PSGL-1, and SCARB2 were detected and compared by immunohistochemistry.
RESULTS:
Strong staining of EV71 -VP1 was observed in the neuron, glial cells, the inflammatory cells of perivascular cuffing, parietal cells of the gastric fundus gland while alveolar macrophages, intestinal gland epithelium cells, mucosa lymphoid nodule and lymphocyte of palatine tonsil showed moderate staining and weak staining were displayed in mesenteric lymph nodes and lymphocyte of spleen. PSGL-1 expression was detected in parietal cells of the gastric fundus gland, tonsillar crypt squamous epithelium, alveolar macrophages and leukocytes in each tissue. SCARB2 expression was observed in all the above tissues except the intestines and spleen.
CONCLUSION
The distribution of EV71 correlates with SCARB2 expression. SCARB2 plays an important role in virus infection and replication. Stomach may be an important site for EV71 replication.
Brain Stem/virology*
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Encephalitis, Viral/virology*
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Enterovirus A, Human/metabolism*
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Enterovirus Infections/virology*
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Humans
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Immunohistochemistry
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Infant
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Leukocytes
;
Lysosome-Associated Membrane Glycoproteins
;
Membrane Glycoproteins/metabolism*
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Receptors, Scavenger/metabolism*
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Receptors, Virus/metabolism*
7.Targeted binding of estradiol with ESR1 promotes proliferation of human chondrocytes by inhibiting activation of ERK signaling pathway.
Min LIU ; Weiwei XIE ; Wei ZHENG ; Danyang YIN ; Rui LUO ; Fengjin GUO
Journal of Southern Medical University 2019;39(2):134-143
OBJECTIVE:
To investigate the effect of estradiol (E2)/estrogen receptor 1 (ESR1) on the proliferation of human chondrocytes and explore the molecular mechanism.
METHODS:
The Ad-Easy adenovirus packaging system was used to construct and package the ESR1-overexpressing adenovirus Ad-ESR1. Western blotting and qPCR were used to detect the expression of ESR1 protein and mRNA in human chondrocyte C28I2 cells. In the cells treated with different adenoviruses, the effects of E2 were tested on the expressions of proteins related with cell autophagy and apoptosis and the phosphorylation of ERK signaling pathway using Western blotting. Immunofluorescence assay was used to observe the intracellular autophagic flow, flow cytometry was performed to analyze the cell apoptosis rate and the cell cycle changes, and qPCR was used to detect the expressions of PCNA, cyclin B1 and cyclin D1 mRNAs. The inhibitory effect of the specific inhibitor of ERK on the expressions of autophagy- and apoptosis-related genes at both the protein and mRNA levels were detected using Western blotting and qPCR.
RESULTS:
Transfection with the recombinant adenovirus overexpressing ESR1 and E2 treatment of C28I2 cells significantly enhanced the expressions of autophagy-related proteins LC3, ATG7, promoted the colocalization of LC3 and LAMP1 in the cytoplasm, increased the expressions of the proliferation-related marker genes PCNA, cyclin B1 and cyclin D1, and supressed the expressions of cleaved caspase-3, caspase-12 and pERK. RNA interference of ESR1 obviously lowered the expression levels of autophagy-related proteins in C28I2 cells, causing also suppression of the autophagic flow, increments of the expressions of apoptosis-related proteins and pERK, and down-regulated the expressions of the proliferation marker genes. Blocking ERK activation with the ERK inhibitor obviously inhibited the effects of E2/ESR1 on autophagy, proliferationrelated gene expressions and cell apoptosis.
CONCLUSIONS
The targeted binding of E2 with ESR1 promotes the proliferation of human chondrocytes possibly by inhibiting the activation of ERK signaling pathway to promote cell autophagy and induce cell apoptosis.
Adenoviridae
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metabolism
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Apoptosis
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Autophagy
;
Autophagy-Related Protein 7
;
metabolism
;
Cell Line
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Cell Proliferation
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Chondrocytes
;
cytology
;
metabolism
;
Estradiol
;
metabolism
;
Estrogen Receptor alpha
;
metabolism
;
Humans
;
Lysosome-Associated Membrane Glycoproteins
;
metabolism
;
MAP Kinase Signaling System
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Microtubule-Associated Proteins
;
metabolism
;
Transfection
8.Relationships between Rapid Eye Movement Sleep Behavior Disorder and Neurodegenerative Diseases: Clinical Assessments, Biomarkers, and Treatment.
Min LI ; Li WANG ; Jiang-Hong LIU ; Shu-Qin ZHAN
Chinese Medical Journal 2018;131(8):966-973
ObjectiveRapid eye movement sleep behavior disorder (RBD) is characterized by dream enactment and loss of muscle atonia during rapid eye movement sleep. RBD is closely related to α-synucleinopathies including Parkinson's disease, dementia with Lewy bodies, and multiple system atrophy. Many studies have investigated the markers of imaging and neurophysiological, genetic, cognitive, autonomic function of RBD and their predictive value for neurodegenerative diseases. This report reviewed the progress of these studies and discussed their limitations and future research directions.
Data SourcesUsing the combined keywords: "RBD", "neurodegenerative disease", "Parkinson disease", and "magnetic resonance imaging", the PubMed/MEDLINE literature search was conducted up to January 1, 2018.
Study SelectionA total of 150 published articles were initially identified citations. Of the 150 articles, 92 articles were selected after further detailed review. This study referred to all the important English literature in full.
ResultsSingle-nucleotide polymorphisms in SCARB2 (rs6812193) and MAPT (rs12185268) were significantly associated with RBD. The olfactory loss, autonomic dysfunction, marked electroencephalogram slowing during both wakefulness and rapid eye movement sleep, and cognitive impairments were potential predictive markers for RBD conversion to neurodegenerative diseases. Traditional structural imaging studies reported relatively inconsistent results, whereas reduced functional connectivity between the left putamen and substantia nigra and dopamine transporter uptake demonstrated by functional imaging techniques were relatively consistent findings.
ConclusionsMore longitudinal studies should be conducted to evaluate the predictive value of biomarkers of RBD. Moreover, because the glucose and dopamine metabolisms are not specific for assessing cognitive cognition, the molecular metabolism directly related to cognition should be investigated. There is a need for more treatment trials to determine the effectiveness of interventions of RBD on preventing the conversion to neurodegenerative diseases.
Biomarkers ; blood ; Humans ; Lysosome-Associated Membrane Glycoproteins ; genetics ; Neurodegenerative Diseases ; blood ; genetics ; physiopathology ; Parkinson Disease ; blood ; genetics ; physiopathology ; Polymorphism, Single Nucleotide ; genetics ; REM Sleep Behavior Disorder ; blood ; genetics ; physiopathology ; Receptors, Scavenger ; genetics ; tau Proteins ; genetics
9.A pH-dependent molecular switch for virion uncoating.
Protein & Cell 2014;5(9):653-654
Capsid Proteins
;
genetics
;
metabolism
;
Enterovirus A, Human
;
genetics
;
metabolism
;
physiology
;
Enterovirus Infections
;
metabolism
;
virology
;
Host-Pathogen Interactions
;
Humans
;
Hydrogen-Ion Concentration
;
Lysosome-Associated Membrane Glycoproteins
;
metabolism
;
RNA, Viral
;
genetics
;
metabolism
;
Receptors, Scavenger
;
metabolism
;
Virion
;
genetics
;
metabolism
;
Virus Attachment
10.LIMP-2 enhances cancer stem-like cell properties by promoting autophagy-induced GSK3β degradation in head and neck squamous cell carcinoma.
Yuantong LIU ; Shujin LI ; Shuo WANG ; Qichao YANG ; Zhizhong WU ; Mengjie ZHANG ; Lei CHEN ; Zhijun SUN
International Journal of Oral Science 2023;15(1):24-24
Cancer stem cell-like cells (CSCs) play an integral role in the heterogeneity, metastasis, and treatment resistance of head and neck squamous cell carcinoma (HNSCC) due to their high tumor initiation capacity and plasticity. Here, we identified a candidate gene named LIMP-2 as a novel therapeutic target regulating HNSCC progression and CSC properties. The high expression of LIMP-2 in HNSCC patients suggested a poor prognosis and potential immunotherapy resistance. Functionally, LIMP-2 can facilitate autolysosome formation to promote autophagic flux. LIMP-2 knockdown inhibits autophagic flux and reduces the tumorigenic ability of HNSCC. Further mechanistic studies suggest that enhanced autophagy helps HNSCC maintain stemness and promotes degradation of GSK3β, which in turn facilitates nuclear translocation of β-catenin and transcription of downstream target genes. In conclusion, this study reveals LIMP-2 as a novel prospective therapeutic target for HNSCC and provides evidence for a link between autophagy, CSC, and immunotherapy resistance.
Humans
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Autophagy
;
Carcinoma, Squamous Cell/pathology*
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Cell Line, Tumor
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Glycogen Synthase Kinase 3 beta/metabolism*
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Head and Neck Neoplasms/pathology*
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Neoplastic Stem Cells/pathology*
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Squamous Cell Carcinoma of Head and Neck/pathology*
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Lysosome-Associated Membrane Glycoproteins