1.Increased expression of PELP1 in human sperm is correlated with decreased semen quality.
Izabela SKIBINSKA ; Miroslaw ANDRUSIEWICZ ; Michal SOIN ; Magdalena JENDRASZAK ; Paulina URBANIAK ; Piotr JEDRZEJCZAK ; Malgorzata KOTWICKA
Asian Journal of Andrology 2018;20(5):425-431
Proline-, glutamic acid-, and leucine-rich protein 1 (PELP1) is a scaffolding protein involved in both genomic and nongenomic estrogen signal transduction pathways. To date, the role of PELP1 protein has yet to be characterized in human sperm and has not been associated with sperm parameters. To confirm the presence of PELP1 in human sperm, fresh semen samples were obtained from 178 donors. The study was designed to establish both mRNA and protein presence, and protein cellular localization. Additionally, the number of PELP1-positive spermatozoa was analyzed in men with normal and abnormal semen parameters. Sperm parameters were assessed according to the World Health Organization (WHO) 2010 standards. The presence of PELP1 in spermatozoa was investigated using four precise, independent techniques. The qualitative presence of transcripts and protein was assessed using reverse transcription-polymerase chain reaction (RT-PCR) and western blot protocols, respectively. The cellular localization of PELP1 was investigated by immunocytochemistry. Quantitative analysis of PELP1-positive cells was done by flow cytometry. PELP1 mRNA and protein was confirmed in spermatozoa. Immunocytochemical analysis identified the presence of PELP1 in the midpieces of human sperm irrespective of sperm parameters. Becton Dickinson fluorescence-activated cell sorting (FACSCalibur™) analysis revealed a significantly lower number of PELP1-positive cells in males with normal semen parameters versus abnormal samples (42.78% ± 11.77% vs 61.05% ± 21.70%, respectively; P = 0.014). The assessment of PELP1 may be a time-saving method used to obtain information about sperm quality. The results of our study suggest that PEPL1 may be utilized as an indicator of sperm quality; thereby, PELP1 may be an additional biomarker useful in the evaluation of male infertility.
Adolescent
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Adult
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Biomarkers/metabolism*
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Co-Repressor Proteins/metabolism*
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Humans
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Infertility, Male/metabolism*
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Male
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Middle Aged
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Semen Analysis
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Sperm Motility/physiology*
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Spermatozoa/metabolism*
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Transcription Factors/metabolism*
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Young Adult
2.Determination of ETO interaction domain within nuclear receptor co-repressor.
Min WANG ; Changlai HAO ; Kejing TANG ; Haiyan XING ; Zheng TIAN ; Yi ZHANG ; Jianxiang WANG
Chinese Journal of Hematology 2002;23(12):621-623
OBJECTIVETo determine the ETO-interaction domain of nuclear receptor co-repressor (N-CoR) for abolishing the biological function of AML1-ETO.
METHODSTen different regions of N-CoR (N-CoRYs) were generated by means of polymerase chain reaction (PCR), and cloned into yeast expression plasmid pGADGL to construct pGADGL/N-CoRYs. The yeast two-hybrid technique and X-gal staining were used to determine the binding between the 10 different regions of N-CoR and ETO.
RESULTSIt was shown that the co-existence of 988-1,126 and 1,551-1,803 amino acid residues of N-CoRY was the ETO-interaction domains required for the binding with ETO.
CONCLUSIONTwo domains of N-CoR that interact with two zinc fingers of ETO, and keep stable binding between the two proteins were identified.
Binding Sites ; genetics ; Humans ; Nuclear Proteins ; chemistry ; genetics ; metabolism ; Nuclear Receptor Co-Repressor 1 ; Plasmids ; genetics ; Protein Binding ; Proto-Oncogene Proteins ; genetics ; metabolism ; RUNX1 Translocation Partner 1 Protein ; Recombinant Fusion Proteins ; genetics ; metabolism ; Repressor Proteins ; chemistry ; genetics ; metabolism ; Transcription Factors ; genetics ; metabolism ; Transfection ; Two-Hybrid System Techniques
3.Inhibitory effect of genistein on the proliferation of Raji cells and its related mechanism.
Jing SHEN ; Jing-Shi WANG ; Li FU ; Da-Yong HUANG ; Zhao WANG
Journal of Experimental Hematology 2014;22(4):971-975
This study was aimed to investigate the anti-proliferative effect of genistein (Gen) on BCL-6 positive Raji cells and its related mechanism. Trypan blue staining and MTT method were used to analyze the anti-proliferative effect of Gen on Raji cells. Cell apoptosis, protein expression and the interaction of BCL-6 and NCoR were detected by PI/AV dual staining, Western blot and Co-IP method, respectively. The results showed that Gen had time- and dose-dependent inhibitory effect on Raji cell proliferation and induced apoptosis. Different dose of Gen had no significant effect on the expression of BCL-6 and NCoR, but could inhibit the binding of BCL-6 and NCoR. It is concluded that Gen shows inhibitory effect on BCL-6 positive lymphoma cells, which can be as a adjuvant therapy for combined rituximab with chemotherapy.
Apoptosis
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drug effects
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Cell Line, Tumor
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Cell Proliferation
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drug effects
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Dose-Response Relationship, Drug
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Genistein
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pharmacology
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Humans
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Lymphocytes
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metabolism
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Nuclear Receptor Co-Repressor 1
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metabolism
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Proto-Oncogene Proteins c-bcl-6
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metabolism
4.Aldolase B attenuates clear cell renal cell carcinoma progression by inhibiting CtBP2.
Mingyue TAN ; Qi PAN ; Qi WU ; Jianfa LI ; Jun WANG
Frontiers of Medicine 2023;17(3):503-517
Aldolase B (ALDOB), a glycolytic enzyme, is uniformly depleted in clear cell renal cell carcinoma (ccRCC) tissues. We previously showed that ALDOB inhibited proliferation through a mechanism independent of its enzymatic activity in ccRCC, but the mechanism was not unequivocally identified. We showed that the corepressor C-terminal-binding protein 2 (CtBP2) is a novel ALDOB-interacting protein in ccRCC. The CtBP2-to-ALDOB expression ratio in clinical samples was correlated with the expression of CtBP2 target genes and was associated with shorter survival. ALDOB inhibited CtBP2-mediated repression of multiple cell cycle inhibitor, proapoptotic, and epithelial marker genes. Furthermore, ALDOB overexpression decreased the proliferation and migration of ccRCC cells in an ALDOB-CtBP2 interaction-dependent manner. Mechanistically, our findings showed that ALDOB recruited acireductone dioxygenase 1, which catalyzes the synthesis of an endogenous inhibitor of CtBP2, 4-methylthio 2-oxobutyric acid. ALDOB functions as a scaffold to bring acireductone dioxygenase and CtBP2 in close proximity to potentiate acireductone dioxygenase-mediated inhibition of CtBP2, and this scaffolding effect was independent of ALDOB enzymatic activity. Moreover, increased ALDOB expression inhibited tumor growth in a xenograft model and decreased lung metastasis in vivo. Our findings reveal that ALDOB is a negative regulator of CtBP2 and inhibits tumor growth and metastasis in ccRCC.
Humans
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Carcinoma, Renal Cell/genetics*
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Fructose-Bisphosphate Aldolase/metabolism*
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Co-Repressor Proteins/metabolism*
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Transcription Factors/genetics*
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Kidney Neoplasms/genetics*
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Cell Line, Tumor
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Cell Proliferation/genetics*
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Gene Expression Regulation, Neoplastic
5.Repressor of GATA-3 can negatively regulate the expression of T cell cytokines through modulation on inducible costimulator.
Yuan-Sheng ZANG ; Zheng FANG ; Yong-An LIU ; Bing LI ; Qing-Yu XIU
Chinese Medical Journal 2012;125(12):2188-2194
BACKGROUNDThe transcription factor, repressor of GATA-3 (ROG), can simultaneously suppress the expression of T helper cells (Th1 and Th2) cytokines. Since the suppression of Th2 cytokines by GATA-3 is well understood, it is postulated that there are other molecular targets of ROG that can suppress the expression of the Th1 cytokines. We hypothesized that ROG might suppress the stimulators of T lymphocyte cytokines such as CD3, CD28, and inducible costimulator (ICOS), or indirectly enhance the expression of cytokine suppressors such as T lymphocyte-associated antigen-4 (CTLA-4) and CD45. The objective of this study was to clarify the molecular targets of ROG involved in suppressing Th1 or Th2 cytokines.
METHODSReal-time quantitative PCR (RT-PCR) and Western blotting were performed to evaluate the mRNA and protein levels of CD3, CD28, ICOS, CTLA-4, and CD45 in Th1 and Th2 cells during various levels of ROG expression. Enzyme-linked immunosorbent assay (ELISA) was used to measure the levels of interferon-γ (IFN-γ) and interleukin (IL)-4 in culture media of Th1 and Th2 cells.
RESULTSThe results showed that the mRNA and protein levels of ROG were relatively low in Th1 and Th2 cells (P < 0.01). After ROG-pcDNA3.1 transfection, the mRNA and protein level of ROG was significantly elevated, while the expression of ICOS, IFN-γ, and IL-4 was markedly down-regulated (P < 0.01). Conversely, transfection of ROG-siRNA led to inhibition of ROG expression and up-regulation of ICOS, IFN-γ and IL-4 (P < 0.01). However, the expression levels of CD3, CD28, CTLA-4 and CD45 did not change in either ROG-pcDNA3.1 or ROG-siRNA-transfected Th1 and Th2 cells (P > 0.05).
CONCLUSIONIt is concluded that ROG can inhibit the expression of Th1 and Th2 cytokines by down-regulating the expression of ICOS, which might be a potential molecular target for asthma treatment.
Animals ; Blotting, Western ; CD28 Antigens ; metabolism ; CD3 Complex ; metabolism ; CD4-Positive T-Lymphocytes ; metabolism ; CTLA-4 Antigen ; metabolism ; Cells, Cultured ; Cytokines ; metabolism ; Enzyme-Linked Immunosorbent Assay ; Inducible T-Cell Co-Stimulator Protein ; metabolism ; Interferon-gamma ; metabolism ; Interleukin-4 ; metabolism ; Leukocyte Common Antigens ; metabolism ; Male ; Mice ; Mice, Inbred BALB C ; Real-Time Polymerase Chain Reaction ; Repressor Proteins ; genetics ; metabolism ; T-Lymphocytes ; metabolism ; Th1 Cells ; metabolism ; Th2 Cells ; metabolism
6.Vitamin C induces periodontal ligament progenitor cell differentiation via activation of ERK pathway mediated by PELP1.
Yan YAN ; Wenfeng ZENG ; Shujun SONG ; Fayun ZHANG ; Wenxi HE ; Wei LIANG ; Zhongying NIU
Protein & Cell 2013;4(8):620-627
The differentiation of periodontal ligament (PDL) progenitor cells is important for maintaining the homeostasis of PDL tissue and alveolar bone. Vitamin C (VC), a water-soluble nutrient that cannot be biosynthesized by humans, is vital for mesenchymal stem cells differentiation and plays an important role in bone remodeling. Therefore, the objective of this study was to determine the function and mechanism of VC in PDL progenitor cells osteogenic differentiation at the molecular level. We demonstrated that VC could induce the osteogenic differentiation and maturation of PDL progenitor cell without other osteogenic agents. During the process, VC preferentially activated ERK1/2 but did not affect JNK or p38. Co-treatment with ERK inhibitor effectively decreased the Vitamin C-induced expression of Runx2. ERK inhibitor also abrogated Vitamin C-induced the minimized nodules formation. PELP1, a nuclear receptor co-regulator, was up-regulated under VC treatment. PELP1 knockdown inhibited ERK phosphorylation. The overexpression of PELP1 had a positive relationship with Runx2 expression. Taken together, we could make a conclude that VC induces the osteogenic differentiation of PDL progenitor cells via PELP1-ERK axis. Our finding implies that VC may have a potential in the regeneration medicine and application to periodontitis treatment.
Ascorbic Acid
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pharmacology
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Butadienes
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pharmacology
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Cell Differentiation
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drug effects
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Cells, Cultured
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Co-Repressor Proteins
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antagonists & inhibitors
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genetics
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metabolism
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Core Binding Factor Alpha 1 Subunit
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genetics
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metabolism
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Humans
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MAP Kinase Signaling System
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drug effects
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Mitogen-Activated Protein Kinase 1
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antagonists & inhibitors
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metabolism
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Mitogen-Activated Protein Kinase 3
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antagonists & inhibitors
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metabolism
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Nitriles
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pharmacology
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Periodontal Ligament
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cytology
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Phosphorylation
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drug effects
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RNA Interference
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RNA, Small Interfering
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metabolism
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Stem Cells
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cytology
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Transcription Factors
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antagonists & inhibitors
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genetics
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metabolism
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Up-Regulation
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drug effects