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LRWD1 Regulates Microtubule Nucleation and Proper Cell Cycle Progression in the Human Testicular Embryonic Carcinoma Cells
Authors:Chia‐Yih Wang  Yu‐Han Hong  Jhih‐Siang Syu  Yung‐Chieh Tsai  Xiu‐Ying Liu  Ting‐Yu Chen  Yin‐Mei Su  Pao‐Lin Kuo  Yung‐Ming Lin  Yen‐Ni Teng
Institution:1. Department of Cell Biology and Anatomy, College of Medicine, National Cheng Kung University, Tainan, Taiwan;2. Institute of Basic Medical Sciences, College of Medicine, National Cheng Kung University, Tainan, Taiwan;3. Department of Biological Sciences and Technology, National University of Tainan, Tainan, Taiwan;4. Department of Obstetrics and Gynecology, Chi‐Mei Medical Center, Tainan, Taiwan;5. Department of Medicine, Taipei Medical University, Taipei, Taiwan;6. Department of Sport Management, Chia Nan University of Pharmacy and Science, Tainan, Taiwan;7. Department of Obstetrics and Gynecology, College of Medicine, National Cheng Kung University, Tainan, Taiwan;8. Department of Urology, College of Medicine, National Cheng Kung University, Tainan, Taiwan
Abstract:Leucine‐rich repeats and WD repeat domain containing protein 1 (LRWD1) is a testis‐specific protein that mainly expressed in the sperm neck where centrosome is located. By using microarray analysis, LRWD1 is identified as a putative gene that involved in spermatogenesis. However, its role in human male germ cell development has not been extensively studied. When checking in the semen of patients with asthenozoospermia, teratozoospermia, and asthenoteratozoospermia, the level of LRWD1 in the sperm neck was significantly reduced with a defective neck or tail. When checking the sub‐cellular localization of LRWD1 in the cells, we found that LRWD1 resided in the centrosome and its centrosomal residency was independent of microtubule transportation in NT2/D1, the human testicular embryonic carcinoma, cell line. Depletion of LRWD1 did not induce centrosome re‐duplication but inhibited microtubule nucleation. In addition, the G1 arrest were observed in LRWD1 deficient NT2/D1 cells. Upon LRWD1 depletion, the levels of cyclin E, A, and phosphorylated CDK2, were reduced. Overexpression of LRWD1 promoted cell proliferation in NT2/D1, HeLa, and 239T cell lines. In addition, we also observed that autophagy was activated in LRWD1 deficient cells and inhibition of autophagy by chloroquine or bafilomycin A1 promoted cell death when LRWD1 was depleted. Thus, we found a novel function of LRWD1 in controlling microtubule nucleation and cell cycle progression in the human testicular embryonic carcinoma cells. J. Cell. Biochem. 119: 314–326, 2018. © 2017 Wiley Periodicals, Inc.
Keywords:LRWD1  CENTROSOME  CELL CYCLE  MICROTUBULE NUCLEATION  AUTOPHAGY
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