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| Establishment and Phenotype Analysis of SHBG Knockout Mouse Model |
| SUN Yi-ping1, WANG Yue1, JIN Zhen1, WANG Xiao-yan1, SUN Lei1, ZHANG Xuan1, FENG Chong1, ZHOU Xiao-hua2 |
1. Department of Obstetrics and Gynecology, Shengjing Hospital, China Medical University, Shenyang 110004, China;
2. Shanghai Research Center for Biomodel Organism, Shanghai 201203, China |
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Abstract Objective:To generate sex hormone binding globulin(SHBG)conditional knockout mice model. In order to investigate the physiological function of SHBG in vivo and to provide experimental means for the study of the relationship between SHBG and gestational diabetes mellitus. Methods:The mouse genomic DNA sequence of SHBG was verified through bioinformatic analysis. According to the SHBG genomic DNA sequence, the gene targeting and knockout vector were constructed. Transfection of the vectors to ES cells by electroporation was performed according to common protocol. Positive ES cells were screened and identified by PCR. Therefore, the dual selected ES cells were microinjected into blastula, then blastula transplantations into the host mice. The chimeric mice were mated with C57BL/6J mice,and the Flox mice were obtained after screening. The Flox mice were hybridized with EⅡA-Cre transgenic mice,and the progeny of the SHBG gene knockout (SHBG-/-) mice were obtained by autocopuation for several times. Results:Several Flox homozygous mice and SHBG gene knockout mice were successfully obtained. Compared with control mice,homozygous mice of SHBG gene knockout were well developed and had reproductive ability. The growth and development of SHBG knockout mice were not significantly different from that of wild type mice.Conclusion:Homozygous mice model of SHBG gene knockout was successfully established,which laid the foundation for further study of the role of SHBG in the gestational diabetes. The SHBG gene knockout mouse model was successfully established and the preliminary phenotypic analysis was performed, which laid the foundation for further study on the role of SHBG in gestational diabetes mellitus. SHBG gene knockout mice were normal in appearance. Due to the limited number of samples and many unknown biological characteristics of gene knockout mice, it needs further study.
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Received: 03 January 2017
Published: 25 August 2017
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[1] Schobert R,Bernhardt G,Biersack B,et al. Steroid conjugates of dichloro (6-aminomethylnicotinate) platinum(Ⅱ):effects on DNA,sex hormone binding globulin,the estrogen receptor,and various breast cancer cell lines. Chem Med Chem,2007,2(3):333-342.
[2] Hammond G L,Bocchinfuso W P. Sex hormone-binding globulin:gene organization and structure/function analyses. Horm Res,1996,45(3-5):197-201.
[3] Hammond G L,Wu T S,Simard M,et al. Evolving utility of sex hormone-binding globulin measurements in clinical medicine. Curr Opin Endocrinol Diabetes Obes,2012,19(3):183-189.
[4] Mehrabian F,Rezae M. Sex hormone binding globulin measurement before conception as a predictor of gestational diabetes in women with polycystic ovarian syndrome. J Res Med Sci,2013,18(8):637-640.
[5] Tong G,Hua X,Zhong Y,et al.Intensive insulin therapy increases sex hormone-binding globulin in newly diagnosed type 2 diabetic patients. Eur J Endocrinol,2014,170(2):237-245.
[6] Wallace I R,McKinley M C,Bell P M,et al. Sex hormone binding globulin and insulin resistance. Clinical Endocrinology,2013,78(3):321-329.
[7] Perry J R,Weedon M N,Langenberg C,et al.Generic evidence that raised sex hormone binding globulin(SHBG)levels reduce the risk of type 2 diabetes. Hum Mol Genet,2010,19(3):535-544.
[8] Ding E L,Song Y,Manson J E,et al. Plasma sex steroid hormones and risk of developing type 2 diabetes in women:a prospective study.Diabetologia,2007,50(10):2076-2084.
[9] Xita N,Tsatsoulis A,Chatzikyriakidou A,et al. Association of the(TAAAA)n repeat polymorphism in the sex hormone-binding globulin (SHBG) gene with polycystic ovary syndrome and relation to SHBG serum levels. J Clin Endocrinol Metab,2003,88(12):5976-5980.
[10] Haiman C A,Riley S E,Freedman M L,et al.Common genetic variation in the sex steroid hormone-binding glogulin (SHBG) gene and circulating shbg levels among postmenopausal women:the Multiethnic Cohort. J Clin Endocrinol Metab,2005,90(4):2198-2204.
[11] 练思宇,金镇,孙磊,等.性激素结合球蛋白基因启动子(TAAAA)n重复多态与妊娠期糖尿病发病的关系.中华妇产科杂志,2014,49(2):138-139. Lian S Y,Jin Z,Sun L,et al. Association of the (TAAAA)n repeat polymorphism of SHBG gene promoter with gestational diabetes mellitus.Chinese Journal of Obstetrics and Gynecology,2014,49(2):138-139.
[12] 息小雪,练思宇,张璇,等.胎盘性激素结合球蛋白基因启动子n重复多态与妊娠期糖尿病关系研究.中国实用妇科与产科杂志,2015,31(12):1142-1145. Xi X X,Lian S Y,Zhang X, et al.Analysis of the relationship between SHBG gene(TAAAA)n polymorphism in placenta and GDM. Chinese Journal of Practical Gynecology and Obstetrics,2015,31(12):1142-1145.
[13] 孙倩,孙磊,金镇,等.RNA干扰技术抑制SHBG基因表达对滋养细胞凋亡的影响.中国医科大学学报,2013,42(8):689-692. Sun Q,Sun L,Jin Z,et al. Effect of siRNA-mediated SHBG gene silencing on apoptosis of trophoblasts.Journal of China Medical University,2013,42(8):689-692.
[14] 范杰慧,金镇,孙倩,等.RNA干扰SHBG基因表达对滋养细胞增殖的影响.中国优生与遗传杂志,2015,23(2):3-7. Fan J H,Jin Z,Sun Q,et al. Effect of RNA silence SHBG gene on proliferation of trophoblasts.Chinese Journal of Birth Health & Heredity,2015,23(2):3-7.
[15] 姜洁璇,孙磊,金镇,等.性激素结合球蛋白和葡萄糖转运蛋白在妊娠期糖尿病胎盘组织中的表达及相关性. 中国医科大学学报,2015,44(1):5-9. Jiang J X,Sun L,Jin Z,et al. Expression of SHBG and GLUT in gestational diabetes placenta tissue and their correlation. Journal of China Medical University,2015,44(1):5-9.
[16] 王越,金镇,孙磊,等. 小鼠性激素结合球蛋白条件基因打靶载体的构建. 中国医科大学学报,2015,44(03):203-208. Wang Y,Jin Z,Sun L,et al. Construction of targeting vector for conditional knockout of murine SHBG. Journal of China Medical University,2015,44(03):203-208. |
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