Effects of cold-inducible RNA-binding protein on the proliferation and apoptosis of spermatogenic cells in vitro following heat stress
Guiping Cheng
A * , Dandan Ke A B * , Meng Rao A C * , Shifu Hu A , Yingying Wang A , Fang Zhou A , Heyu Liu A , Changhong Zhu A D and Wei Xia A D E
+ Author Affiliations
- Author Affiliations
A Family Planning Research Institute, Tongji Medical College, Huazhong University of Science and Technology, No. 13 Hang-kong Road, Wuhan 430030, China.
B Department of Obstetrics and Gynecological Ultrasound Imaging, Renmin Hospital of Wuhan University, No. 99 Zhang zhidong Road, No. 238 Jiefang Road, Wuhan 430060, China.
C Department of Reproduction and Genetics, The First Affiliated Hospital of Kunming Medical University, No. 295 Xichang Road, Kunming 650032, China.
D Reproductive Medicine Center, Tongji Medical College, Huazhong University of Science and Technology, No. 128 Sanyang Road, Wuhan 430010, China.
E Corresponding author. Email: tjxiawei@hust.edu.cn
Reproduction, Fertility and Development 31(5) 953-961 https://doi.org/10.1071/RD18469
Submitted: 30 August 2018 Accepted: 21 December 2018 Published: 12 February 2019
Abstract
Cold-inducible RNA-binding protein (CIRBP) is reduced by scrotal hyperthermia in cryptorchidism, varicocoele and heat treatment, but there is no direct evidence clarifying the relationship between CIRBP and spermatogenesis. The aim of this study was to investigate the expression of CIRBP in GC2-spd cells (a mouse spermatocyte cell line) before and after heat treatment, and to determine the effects of the downregulation or overexpression of CIRBP on spermatocyte cell proliferation and apoptosis. GC2-spd cells overexpressing CIRBP and GC2-spd cells in CIRBP was knocked down were constructed to investigate the function of CIRBP in cell proliferation and apoptosis using a cell counting kit-8 and flow cytometry respectively. In addition, proliferation and apoptosis were evaluated in GC2-spd cells that had been heated for 30 or 60 min, and were analysed 12, 24, and 48 h after heat treatment. Heat treatment clearly suppressed the proliferation of GC2-spd cells, and upregulation of CIRBP expression in GC2-spd cells promoted cell proliferation and decreased apoptosis before and after heat stress; in contrast, downregulation of CIRBP expression inhibited cell proliferation and increased apoptosis. These findings suggest that CIRBP exerts a protective effect against spermatogenic injury caused by heat stress.
Additional keywords: CIRBP, spermatocytes.
References
Absalan, F., Movahedin, M., and Mowla, S. J. (2012). Evaluation of apoptotic genes expression and its protein after treatment of cryptorchid mice. Iran. Biomed. J. 16, 77–83.| 22801280PubMed |
Banks, S., King, S. A., Irvine, D. S., and Saunders, P. T. K. (2005). Impact of a mild scrotal heat stress on DNA integrity in murine spermatozoa. Reproduction 129, 505–514.
| Impact of a mild scrotal heat stress on DNA integrity in murine spermatozoa.Crossref | GoogleScholarGoogle Scholar | 15798026PubMed |
Burd, C. G., and Dreyfuss, G. (1994). Conserved structures and diversity of functions of RNA-binding proteins. Science 265, 615–621.
| Conserved structures and diversity of functions of RNA-binding proteins.Crossref | GoogleScholarGoogle Scholar | 8036511PubMed |
Chen, M., Yuan, J., Shi, Y., Zhang, X., Hu, Z., Gao, F., and Liu, Y. (2008). Effect of 43 degrees treatment on expression of heat shock proteins 105, 70 and 60 in cultured monkey Sertoli cells. Asian J. Androl. 10, 474–485.
| Effect of 43 degrees treatment on expression of heat shock proteins 105, 70 and 60 in cultured monkey Sertoli cells.Crossref | GoogleScholarGoogle Scholar | 18385910PubMed |
De Leeuw, F., Zhang, T., Wauquier, C., Huez, G., Kruys, V., and Gueydan, C. (2007). The cold-inducible RNA-binding protein migrates from the nucleus to cytoplasmic stress granules by a methylation-dependent mechanism and acts as a translational repressor. Exp. Cell Res. 313, 4130–4144.
| The cold-inducible RNA-binding protein migrates from the nucleus to cytoplasmic stress granules by a methylation-dependent mechanism and acts as a translational repressor.Crossref | GoogleScholarGoogle Scholar | 17967451PubMed |
Fornace, A. J. J., Alamo, I. J., and Hollander, M. C. (1988). DNA damage-inducible transcripts in mammalian cells. Proc. Natl Acad. Sci. USA 85, 8800–8804.
| DNA damage-inducible transcripts in mammalian cells.Crossref | GoogleScholarGoogle Scholar |
Hikim, A. P. S., Lue, Y., Yamamoto, C. M., Vera, Y., Rodriguez, S., Yen, P. H., Soeng, K., Wang, C., and Swerdloff, R. S. (2003). Key apoptotic pathways for heat-induced programmed germ cell death in the testis. Endocrinology 144, 3167–3175.
| Key apoptotic pathways for heat-induced programmed germ cell death in the testis.Crossref | GoogleScholarGoogle Scholar |
Jannes, P., Spiessens, C., Van der Auwera, I., D’Hooghe, T., Verhoeven, G., and Vanderschueren, D. (1998). Male subfertility induced by acute scrotal heating affects embryo quality in normal female mice. Hum. Reprod. 13, 372–375.
| Male subfertility induced by acute scrotal heating affects embryo quality in normal female mice.Crossref | GoogleScholarGoogle Scholar | 9557841PubMed |
Jian, F., Chen, Y., Ning, G., Fu, W., Tang, H., Chen, X., Zhao, Y., Zheng, L., Pan, S., Wang, W., Bian, L., and Sun, Q. (2016). Cold inducible RNA binding protein upregulation in pituitary corticotroph adenoma induces corticotroph cell proliferation via Erk signaling pathway. Oncotarget 7, 9175–9187.
| Cold inducible RNA binding protein upregulation in pituitary corticotroph adenoma induces corticotroph cell proliferation via Erk signaling pathway.Crossref | GoogleScholarGoogle Scholar | 26824322PubMed |
Kheradmand, A., Dezfoulian, O., and Alirezaei, M. (2012). Ghrelin regulates Bax and PCNA but not Bcl-2 expressions following scrotal hyperthermia in the rat. Tissue Cell 44, 308–315.
| Ghrelin regulates Bax and PCNA but not Bcl-2 expressions following scrotal hyperthermia in the rat.Crossref | GoogleScholarGoogle Scholar | 22658447PubMed |
Liu, J., Xue, J., Zhang, H., Li, S., Liu, Y., Xu, D., Zou, M., Zhang, Z., and Diao, J. (2015). Cloning, expression, and purification of cold inducible RNA-binding protein and its neuroprotective mechanism of action. Brain Res. 1597, 189–195.
| Cloning, expression, and purification of cold inducible RNA-binding protein and its neuroprotective mechanism of action.Crossref | GoogleScholarGoogle Scholar | 25498861PubMed |
Lue, Y. H., Hikim, A. P., Swerdloff, R. S., Im, P., Taing, K. S., Bui, T., Leung, A., and Wang, C. (1999). Single exposure to heat induces stage-specific germ cell apoptosis in rats: role of intratesticular testosterone on stage specificity. Endocrinology 140, 1709–1717.
| Single exposure to heat induces stage-specific germ cell apoptosis in rats: role of intratesticular testosterone on stage specificity.Crossref | GoogleScholarGoogle Scholar | 10098507PubMed |
Lue, Y., Hikim, A. P., Wang, C., Im, M., Leung, A., and Swerdloff, R. S. (2000). Testicular heat exposure enhances the suppression of spermatogenesis by testosterone in rats: the ‘two-hit’ approach to male contraceptive development. Endocrinology 141, 1414–1424.
| Testicular heat exposure enhances the suppression of spermatogenesis by testosterone in rats: the ‘two-hit’ approach to male contraceptive development.Crossref | GoogleScholarGoogle Scholar | 10746645PubMed |
Mieusset, R., Bujan, L., Mondinat, C., Mansat, A., Pontonnier, F., and Grandjean, H. (1987). Association of scrotal hyperthermia with impaired spermatogenesis in infertile men. Fertil. Steril. 48, 1006–1011.
| Association of scrotal hyperthermia with impaired spermatogenesis in infertile men.Crossref | GoogleScholarGoogle Scholar | 3678498PubMed |
Nishiyama, H., Higashitsuji, H., Yokoi, H., Itoh, K., Danno, S., Matsuda, T., and Fujita, J. (1997). Cloning and characterization of human CIRP (cold-inducible RNA-binding protein) cDNA and chromosomal assignment of the gene. Gene 204, 115–120.
| Cloning and characterization of human CIRP (cold-inducible RNA-binding protein) cDNA and chromosomal assignment of the gene.Crossref | GoogleScholarGoogle Scholar | 9434172PubMed |
Nishiyama, H., Danno, S., Kaneko, Y., Itoh, K., Yokoi, H., Fukumoto, M., Okuno, H., Millán, J. L., Matsuda, T., Yoshida, O., and Fujita, J. (1998). Decreased expression of cold-inducible RNA-binding protein (CIRP) in male germ cells at elevated temperature. Am. J. Pathol. 152, 289–296.
| 9422546PubMed |
Rao, M., Zhao, X., Yang, J., Hu, S., Lei, H., Xia, W., and Zhu, C. (2015). Effect of transient scrotal hyperthermia on sperm parameters, seminal plasma biochemical markers, and oxidative stress in men. Asian J. Androl. 17, 668–675.
| Effect of transient scrotal hyperthermia on sperm parameters, seminal plasma biochemical markers, and oxidative stress in men.Crossref | GoogleScholarGoogle Scholar | 25652627PubMed |
Rao, M., Xia, W., Yang, J., Hu, L., Hu, S., Lei, H., Wu, Y., and Zhu, C. (2016). Transient scrotal hyperthermia affects human sperm DNA integrity, sperm apoptosis, and sperm protein expression. Andrology 4, 1054–1063.
| Transient scrotal hyperthermia affects human sperm DNA integrity, sperm apoptosis, and sperm protein expression.Crossref | GoogleScholarGoogle Scholar | 27410176PubMed |
Rao, M., Ma, S., Hu, S., Lei, H., Wu, Y., Zhou, Y., Xia, W., and Zhu, C. (2017). Isobaric tags for relative and absolute quantification-based proteomic analysis of testis biopsies in rhesus monkeys treated with transient scrotal hyperthermia. Oncotarget 8, 85909–85925.
| Isobaric tags for relative and absolute quantification-based proteomic analysis of testis biopsies in rhesus monkeys treated with transient scrotal hyperthermia.Crossref | GoogleScholarGoogle Scholar | 29156766PubMed |
Rao, M., Ke, D., Cheng, G., Hu, S., Wu, Y., Wang, Y., Zhou, F., Liu, H., Zhu, C., and Xia, W. (2018). The regulation of CIRBP by transforming growth factor beta during heat shock-induced testicular injury. Andrology , .
| The regulation of CIRBP by transforming growth factor beta during heat shock-induced testicular injury.Crossref | GoogleScholarGoogle Scholar | 30461215PubMed |
Rockett, J. C., Mapp, F. L., Garges, J. B., Luft, J. C., Mori, C., and Dix, D. J. (2001). Effects of hyperthermia on spermatogenesis, apoptosis, gene expression, and fertility in adult male mice. Biol. Reprod. 65, 229–239.
| Effects of hyperthermia on spermatogenesis, apoptosis, gene expression, and fertility in adult male mice.Crossref | GoogleScholarGoogle Scholar | 11420244PubMed |
Saito, K., Fukuda, N., Matsumoto, T., Iribe, Y., Tsunemi, A., Kazama, T., Yoshida-Noro, C., and Hayashi, N. (2010). Moderate low temperature preserves the stemness of neural stem cells and suppresses apoptosis of the cells via activation of the cold-inducible RNA binding protein. Brain Res. 1358, 20–29.
| Moderate low temperature preserves the stemness of neural stem cells and suppresses apoptosis of the cells via activation of the cold-inducible RNA binding protein.Crossref | GoogleScholarGoogle Scholar | 20735994PubMed |
Sakurai, T., Itoh, K., Higashitsuji, H., Nonoguchi, K., Liu, Y., Watanabe, H., Nakano, T., Fukumoto, M., Chiba, T., and Fujita, J. (2006). Cirp protects against tumor necrosis factor-α-induced apoptosis via activation of extracellular signal-regulated kinase. Biochim. Biophys. Acta 1763, 290–295.
| Cirp protects against tumor necrosis factor-α-induced apoptosis via activation of extracellular signal-regulated kinase.Crossref | GoogleScholarGoogle Scholar | 16569452PubMed |
Sheikh, M. S., Carrier, F., Papathanasiou, M. A., Hollander, M. C., Zhan, Q., Yu, K., and Fornace, A. J. J. (1997). Identification of several human homologs of hamster DNA damage-inducible transcripts. Cloning and characterization of a novel UV-inducible cDNA that codes for a putative RNA-binding protein. J. Biol. Chem. 272, 26720–26726.
| Identification of several human homologs of hamster DNA damage-inducible transcripts. Cloning and characterization of a novel UV-inducible cDNA that codes for a putative RNA-binding protein.Crossref | GoogleScholarGoogle Scholar | 9334257PubMed |
Tang, C., Wang, Y., Lan, D., Feng, X., Zhu, X., Nie, P., and Yue, H. (2015). Analysis of gene expression profiles reveals the regulatory network of cold-inducible RNA-binding protein mediating the growth of BHK-21 cells. Cell Biol. Int. 39, 678–689.
| Analysis of gene expression profiles reveals the regulatory network of cold-inducible RNA-binding protein mediating the growth of BHK-21 cells.Crossref | GoogleScholarGoogle Scholar | 25597958PubMed |
Wellmann, S., Buhrer, C., Moderegger, E., Zelmer, A., Kirschner, R., Koehne, P., Fujita, J., and Seeger, K. (2004). Oxygen-regulated expression of the RNA-binding proteins RBM3 and CIRP by a HIF-1-independent mechanism. J. Cell Sci. 117, 1785–1794.
| Oxygen-regulated expression of the RNA-binding proteins RBM3 and CIRP by a HIF-1-independent mechanism.Crossref | GoogleScholarGoogle Scholar | 15075239PubMed |
Xia, Z.-P., Zheng, X.-M., Zheng, H., Liu, X.-J., Liu, G.-Y., and Wang, X.-H. (2012a). Downregulation of cold-inducible RNA-binding protein activates mitogen-activated protein kinases and impairs spermatogenic function in mouse testes. Asian J. Androl. 14, 884–889.
| Downregulation of cold-inducible RNA-binding protein activates mitogen-activated protein kinases and impairs spermatogenic function in mouse testes.Crossref | GoogleScholarGoogle Scholar | 23001445PubMed |
Xia, Z., Zheng, X., Zheng, H., Liu, X., Yang, Z., and Wang, X. (2012b). Cold-inducible RNA-binding protein (CIRP) regulates target mRNA stabilization in the mouse testis. FEBS Lett. 586, 3299–3308.
| Cold-inducible RNA-binding protein (CIRP) regulates target mRNA stabilization in the mouse testis.Crossref | GoogleScholarGoogle Scholar | 22819822PubMed |
Xia, Z., Jiang, K., Liu, T., Zheng, H., Liu, X., and Zheng, X. (2013). The protective effect of cold-inducible RNA-binding protein (CIRP) on testicular torsion/detorsion: an experimental study in mice. J. Pediatr. Surg. 48, 2140–2147.
| The protective effect of cold-inducible RNA-binding protein (CIRP) on testicular torsion/detorsion: an experimental study in mice.Crossref | GoogleScholarGoogle Scholar | 24094970PubMed |
Yamamoto, C. M., Sinha Hikim, A. P., Huynh, P. N., Shapiro, B., Lue, Y., Salameh, W. A., Wang, C., and Swerdloff, R. S. (2000). Redistribution of Bax is an early step in an apoptotic pathway leading to germ cell death in rats, triggered by mild testicular hyperthermia. Biol. Reprod. 63, 1683–1690.
| Redistribution of Bax is an early step in an apoptotic pathway leading to germ cell death in rats, triggered by mild testicular hyperthermia.Crossref | GoogleScholarGoogle Scholar | 11090436PubMed |
Yang, C., and Carrier, F. (2001). The UV-inducible RNA-binding protein A18 (A18 hnRNP) plays a protective role in the genotoxic stress response. J. Biol. Chem. 276, 47277–47284.
| The UV-inducible RNA-binding protein A18 (A18 hnRNP) plays a protective role in the genotoxic stress response.Crossref | GoogleScholarGoogle Scholar | 11574538PubMed |
Yin, Y., Hawkins, K. L., DeWolf, W. C., and Morgentaler, A. (1997). Heat stress causes testicular germ cell apoptosis in adult mice. J. Androl. 18, 159–165.
| 9154510PubMed |
Yuan, J.-L., Zhang, Y.-T., and Wang, Y. (2010). Increased apoptosis of spermatogenic cells in cryptorchidism rat model and its correlation with transforming growth factor beta type II receptor. Urology 75, 992–998.
| Increased apoptosis of spermatogenic cells in cryptorchidism rat model and its correlation with transforming growth factor beta type II receptor.Crossref | GoogleScholarGoogle Scholar | 19931125PubMed |
Zeng, Y., Kulkarni, P., Inoue, T., and Getzenberg, R. H. (2009). Down-regulating cold shock protein genes impairs cancer cell survival and enhances chemosensitivity. J. Cell. Biochem. 107, 179–188.
| Down-regulating cold shock protein genes impairs cancer cell survival and enhances chemosensitivity.Crossref | GoogleScholarGoogle Scholar | 19277990PubMed |
Zhang, K., Shang, Y., Liao, S., Zhang, W., Nian, H., Liu, Y., Chen, Q., and Han, C. (2007). Uncoupling protein 2 protects testicular germ cells from hyperthermia-induced apoptosis. Biochem. Biophys. Res. Commun. 360, 327–332.
| Uncoupling protein 2 protects testicular germ cells from hyperthermia-induced apoptosis.Crossref | GoogleScholarGoogle Scholar | 17603020PubMed |
Zhang, H.-T., Xue, J.-H., Zhang, Z.-W., Kong, H.-B., Liu, A.-J., Li, S.-C., and Xu, D.-G. (2015). Cold-inducible RNA-binding protein inhibits neuron apoptosis through the suppression of mitochondrial apoptosis. Brain Res. 1622, 474–483.
| Cold-inducible RNA-binding protein inhibits neuron apoptosis through the suppression of mitochondrial apoptosis.Crossref | GoogleScholarGoogle Scholar | 26168889PubMed |
Zhou, K.-W., Zheng, X.-M., Yang, Z.-W., Zhang, L., and Chen, H.-D. (2009). Overexpression of CIRP may reduce testicular damage induced by cryptorchidism. Clin. Invest. Med. 32, E103–E111.
| Overexpression of CIRP may reduce testicular damage induced by cryptorchidism.Crossref | GoogleScholarGoogle Scholar | 19331798PubMed |
Zhu, H., Cui, Y., Xie, J., Chen, L., Chen, X., Guo, X., Zhu, Y., Wang, X., Tong, J., Zhou, Z., Jia, Y., Lue, Y. H., Hikim, A. S., Wang, C., Swerdloff, R. S., and Sha, J. (2010). Proteomic analysis of testis biopsies in men treated with transient scrotal hyperthermia reveals the potential targets for contraceptive development. Proteomics 10, 3480–3493.
| Proteomic analysis of testis biopsies in men treated with transient scrotal hyperthermia reveals the potential targets for contraceptive development.Crossref | GoogleScholarGoogle Scholar | 20815088PubMed |
