肿瘤抑制蛋白基因Tp53在牦牛体外受精早期胚胎中的表达

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摘要本研究旨在探讨肿瘤抑制蛋白基因(tumor protein p53, Tp53)在牦牛(Bos grunneins)早期不同发育时期体外受精胚胎中的表达情况及其与体外受精胚胎发育的关系。本实验采集牦牛卵巢，收集牦牛卵丘-卵母细胞复合体(cumulus-oocyte complexes, COCs)进行体外成熟培养及对成熟卵母细胞体外受精后进行培养。采用RT-PCR、实时荧光定量PCR(quantitative Real-time PCR, qRT-PCR)和间接免疫荧光方法检测不同时期体外受精胚胎发育过程中Tp53在基因和蛋白水平的表达动态。结果表明，在不同发育时期体外受精胚胎中均有Tp53基因的表达，并且Tp53在2~4细胞期和囊胚期表达量相对较低，组间差异不显著(P＞0.05)；Tp53在5~8细胞期和9~16期表达量相对较高，组间差异不显著(P＞0.05)；在2~4细胞期和囊胚期表达量与在4~8细胞期和9~16期表达量相比差异显著(P＜0.05)。本研究表明，Tp53在调节牦牛早期胚胎发育中也起着非常重要的作用。Tp53的表达量与胚胎发育阻断的发生几率成正相关。间接免疫荧光显示Tp53蛋白主要存在于细胞核中，少量存在于细胞质中，其作用和DNA的损伤有关。

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	郑红飞
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	张译夫
	吕鹏
	崔燕
	余四九

关键词 ：牦牛, 体外受精, 胚胎, 肿瘤抑制蛋白基因Tp53

Abstract：The objective of this study was to research the expression of tumor protein p53 (Tp53) in yak (Bos grunneins) during different development period of embryo and to verify the relationship with in vitro fertilization (IVF) embryo development. In this study, we gathered the ovarian of female yak and collected cumulus-oocyte complexes (COCs) during in vitro maturation and to culture these IVF embryos. The expression of Tp53 in different periods of IVF embryos were detected by the methods of PCR, quantitative Real-time PCR and immunofluorescence at mRNA and protein levels. The results showed that Tp53 gene expressed in different development period of IVF embryos, and Tp53 in blastocysts and 2~4 cells volume was relatively low, and there was no significant difference between groups (P＞0.05); Tp53 in stage 5~8 cells and 9~16 expression quantity was relatively high, and there was no significant difference between groups (P＞0.05). The expression of Tp53 in stage 2~4 cells and blastocyst had significant difference compared with 5~8 cell embryo and 9~16 cell embryo(P＜0.05). This study suggests that Tp53 played a very important role in adjusting early embryonic development. The relationship between the expression of Tp53 and developmental block was positive correlation. Indirect immune fluorescent displayed that Tp53 mainly existed in the nucleus while a small amount existed in cytoplasm and its role was related to DNA damage.

Key words： Bos grunniens In vitro fertilization (IVF) Embryo Tumor protein p53 gene (Tp53)

收稿日期: 2015-02-02 出版日期: 2015-07-09

基金资助:牦牛体内胚胎、体外受精胚胎和核移植胚胎附植前细胞凋亡的特征及其机理;牦牛生长激素受体、促卵泡素受体和促黄体素受体基因特性的研究与应用

通讯作者: 余四九 E-mail: sjyu@163.com

引用本文:

郑红飞潘阳阳李秦张译夫吕鹏崔燕余四九. 肿瘤抑制蛋白基因Tp53在牦牛体外受精早期胚胎中的表达[J]. , 2015, 23(9): 1240-1245.

链接本文:

http://journal05.magtech.org.cn/Jwk_ny/CN/ 或 http://journal05.magtech.org.cn/Jwk_ny/CN/Y2015/V23/I9/1240

Levine A J, Oren M. 2009. The first 30 years of p53: growing ever more complex[J]. Nature Reviews Cancer, 9(10): 749-758.
Lozano G, Zambetti G P. 2005. What have animal models taught us about the p53 pathway?[J]. The Journal of pathology, 205(2): 206-220.
Yap D B S, Hsieh J K, Zhong S, et al. 2004. Ser392 phosphorylation regulates the oncogenic function of mutant p53[J]. Cancer research, 64(14): 4749-4754.
Fridman J S, Lowe S W. 2003.Control of apoptosis by p53[J]. Oncogene, 22(56): 9030-9040.
Hu W. 2009.The role of p53 gene family in reproduction[J]. Cold Spring Harbor perspectives in biology, 1(6): a001073.
Vogelstein B, Lane D, Levine A J. 2000. Surfing the p53 network[J]. Nature, 408(6810): 307-310.
Torchinsky A, Toder V. 2010. Mechanisms of the embryo's response to embryopathic stressors: a focus on p53[J]. Journal of reproductive immunology, 85(1): 76-80.
李拥军, 敖红, 孙桂金, 等. 2005. 体外培养山羊早期胚胎基因表达的研究[J]. 扬州大学学报: 农业与生命科学版, 26(1): 37-40. (Li Y J,Ao H,Sun GJ,ey al.2005.Preliminary study on the gene expression of preimplantation goat embryos produced in vitro[J]. Journal of yangzhou university: Agriculture and life sciences, 26(1): 37-40.)
Hussein M R. 2005. Apoptosis in the ovary: molecular mechanisms[J]. Human reproduction update, 11(2): 162-178.
Urist M, Prives C. 2002. p53 leans on its siblings[J]. Cancer Cell, 1(4): 311-313.
Hale T K, Braithwaite A W. 1995. Identification of an upstream region of the mouse p53 promoter critical for transcriptional expression[J]. Nucleic acids research, 23(4): 663-669.
Shaw P, Bovey R, Tardy S, et al. 1992. Induction of apoptosis by wild-type p53 in a human colon tumor-derived cell line[J]. Proceedings of the National Academy of Sciences, 89(10): 4495-4499.
李拥军. 2005. 山羊早期胚胎发育基因表达的研究 [D][D]. 博士学位论文, 甘肃农业大学,导师：赵有璋.pp.14-25.(Li Y J.2005.Studies on developmental gene expression of goat preimplantation embryos [D][D]. Thesis for Dr. Gansu agricultural university, Supervisor:Zhao Y Z, pp.14-25.)
张进隆. 2008. 奶牛卵母细胞及早期胚胎基因表达分析[D]. 硕士学位论文，甘肃农业大学，导师：赵兴绪.pp.68. (Zhang J L.2008.Gene expression analysis of bovine oocytes and preimpletation embryons[D]. Thesis for M.S. Gansu agricultural university ,Supervisor:Zhao X X, pp.68.)
刘爱华. 2007. p53 和 p38 对 UV 诱导的细胞凋亡的抑制效应及其机制研究[D]. 博士学位论文, 第一军医大学，导师：罗深秋. pp.33-34,47-48. (Liu A H.2007.Studies on the Mechanism and the Inhibitory Effects of p53 and p38 MAP kinase on Cell Apoptosis induced by Ultraviolct[D]. Thesis for Dr. The first military medical university，Supervisor: Luo S Q, pp.33-34，47-48.)
Fronza G, Inga A, Monti P, et al. 2000.The yeast< i> p53 functional assay: a new tool for molecular epidemiology. Hopes and facts[J]. Mutation Research/Reviews in Mutation Research, 462(2): 293-301.
Gostissa M, Hofmann T G, Will H, et al. 2003. Regulation of p53 functions: let’s meet at the nuclear bodies[J]. Current opinion in cell biology, 15(3): 351-357.
Pietsch E C, Humbey O, Murphy M E. 2006. Polymorphisms in the p53 pathway[J]. Oncogene, 25(11): 1602-1611.