ASCL2基因在体细胞克隆牛和自然繁殖牛肺脏中的甲基化分析

Abstract

Abstract: In an effort to determine the reprogramming of ASCL2 gene in SCNT bovines，the bisulfite sequencing method （BSP） was used to analyze the DNA methylation status in promoter region of ASCL2 gene in lung tissues of SCNT cattle and natural breeding cattle.The results showed that the methylation level of ASCL2 promoter exhibited seriously hypermethylated in SCNT bovines （28.41%） compared with naturally breed bovines （7.62%）（P＜0.05）. Our results suggested that the abnormal DNA methylation of ASCL2 might be associated with the defects of lung organs in died SCNT bovines，which was one of the main reasons for neonatal death of SCNT animals.

Key words: ASCL2; reprogramming; SCNT bovine; BSP; DNA methylation; organ defect

WANG Meng-nan，ZHANG Ming-yue，LI Ning，LI Shi-jie. DNA Methylation Status of ASCL2 Gene in Lungs of SCNT Cattle and Naturally Reproduced Cattle[J]. China Animal Husbandry & Veterinary Medicine.

References

1 赵丽霞，赵高平，郭荣启，等.绵羊Cdkn1c的DNA甲基化分析［J］.中国畜牧兽医，2010，37（4）：85～88.
2 Arnold D R，Lefebvre R，Smith L C.Characterization of the placenta specific bovine mammalian achaete scute-like homologue 2 （Mash2） gene［J］.Placenta，2006，27(11～12)：1124～1131.
3 Cui X S，Zhang D X，Ko Y G，et al.Aberrant epigenetic reprogramming of imprinted microRNA-127 and Rtl1 in cloned mouse embryos［J］.Biochem Biophys Res Commun，2009，379（2）：390～394.
4 Daniels R，Hall V，Trounson A O.Analysis of gene transcription in bovine nuclear transfer embryos reconstructed with granulosa cell nuclei［J］.Biol Reprod，2000，63（4）：1034～1040.
5 Farin C E，Farin P W，Piedrahita J A.Development of fetuses from in vitro-produced and cloned bovine embryos［J］.J Anim Sci，2004，82：E53～E62.
6 Gall L，Brochard V，Ruffini S，et al.Intermediate filaments promote nuclear mechanical constraints during somatic cell nuclear transfer in the mouse［J］.Cell Reprogram，2012，14（6）：497～504.
7 Gong G，Dai Y，Fan B，et al.Production of transgenic blastocyst by nuclear transfer from different types of somatic cells in cattle［J］.Sci China C Life Sci，2004，47（2）：183～189.
8 Higashimoto K，Soejima H，Saito T，et al.Imprinting disruption of the Cdkn1c/Kcnq1ot1 domain：The molecular mechanisms causing beckwith-wiedemann syndrome and cancer［J］.Cytogenet Genome Res，2006，113(1～4)：306～312.
9 Imamura T，Kerjean A，Heams T，et al.Dynamic CpG and non-CpG methylation of the Peg1/Mest gene in the mouse oocyte and preimplantation embryo［J］.J Biol Chem，2005，280（20）：20171～20175.
10 Kawahara M，Kono T.Roles of genes regulated by two paternally methylated imprinted regions on chromosomes 7 and 12 in mouse ontogeny［J］.J Reprod Dev，2012，58（2）：175～179.
11 Khatib H，Zaitoun I，Kim E S.Comparative analysis of sequence characteristics of imprinted genes in human，mouse and cattle［J］.Mamm Genome，2007，18(6～7)：538～547.
12 Kremenskoy M，Kremenska Y，Suzuki M，et al.Epigenetic characterization of the CpG islands of bovine leptin and POU5F1 genes in cloned bovine fetuses［J］.J Repord Dev，2006，52（2）：277～285.
13 Kwon O H，Park J L，Baek S J，et al.Aberrant upregulation of ASCL2 by promoter demethylation promotes the growth and resistance to 5-fluorouracil of gastric cancer cells［J］.Cancer Science，2013，104（3）：391～397.
14 Li S，Li Y，Du W，et al.Aberrant gene expression in organs of bovine clones that die within two days after birth［J］.Biol Reprod，2005，72（2）：258～265.
15 Lie D C，Colamarino S A，Song H J，et al.Wnt signalling regulates adult hippocampal neurogenesis［J］.Nature，2005，437(7063)：1370～1375.
16 Lorincz M C，Dickerson D R，Schmitt M，et al.Intragenic DNA methylation alters chromatin structure and elongation efficiency in mammalian cells［J］.Nat Struct Mol Biol，2004，11(11)：1068～1075.
17 Lucifero D，Suzuki J，Bordiqnon V，et al.Bovine snrpn methylation imprint in oocytes and day 17 in vitro-produced and somatic cell nuclear transfer embryos［J］.Biol Reprod，2006，75（4）：531～538.
18 Oh-McGinnis R，Bogutz A B，Lefebvre L.Partial loss of ASCL2 function affects all three layers of the mature placenta and causes intrauterine growth restriction［J］.Developmental Biology，2011，351（2）：277～286.
19 Rauch T A，Rauch T A，Wu X，et al.A human B cell methylome at 100-base pair resolution［J］.Proc Natl Acad Sci USA，2009，106（3）：671～678.
20 Reik W，Walter J.Genomic imprinting：Parental influence on the genome［J］.Nat Rev Genet，2001，2（1）：21～32.
21 Rentsendorj A，Mohan S，Szabó P，et al.A genomic imprinting defect in mice traced to a single gene［J］.Genetics，2010，186（3）：917～927.
22 Shen C J，Lin C C，Shen P C，et al.Imprinted genes and satellite loci are differentially methylated in bovine somatic cell nuclear transfer clones［J］.Cell Reprogram，2013，15（5）：413～424.
23 Shiota K，Kogo Y，Ohgane J，et al.Epigenetic marks by DNA methylation specific to stem，germ and somatic cells in mice［J］.Genes Cells，2002，7（9）：961～969.
24 Stepan H，Marqwardt W，Kuhn Y，et al.Structure and regulation of the murine Mash2 gene［J］.Biol Reprod，2003，68（1）：40～44.
25 Umlauf D，Goto Y，Cao R，et al.Imprinting along the Kcnq1 domain on mouse chromosome 7 involves repressive histone methylation and recruitment of polycomb group complexes［J］.Nat Genet，2004，36（12）：1296～1300.
26 Walter J，Paulsen M.Imprinting and disease［J］.Semin Cell Dev Biol，2003，14（1）：101～110.
27 Wei Y，Zhu J，Huan Y，et al.Aberrant expression and methylation status of putatively imprinted genes in placenta of cloned piglets［J］.Cell Reprogram，2010，12（2）：213～222.
28 Wolffe A P.Transcriptional control：Imprinting insulation［J］.Curr Biol，2000，10（12）：463～465.
29 Woods G L，White K L，Vanderwall D K，et al.A mule cloned from fetal cells by nuclear transfer［J］.Science，2003，301（5636）：1063.
30 Xin Z，Allis C D，Wagstaff J.Parent-specific complementary patterns of histone H3 lysine 9 and H3 lysine 4 methylation at the Prader-Willi syndrome imprinting center［J］.Am J Hum Genet，2001，69（6）：1389～1394.
31 Young L E，Fairburn H R.Improving the safety of embryo technologies: Possible role of genomic imprinting［J］.Theriogenology，2000，53（2）：627～648.
32 Zhang D，Yoon H G，Wong J.JMJD2A is a novel N-CoR-interacting protein and is involved in repression of the human transcription factor achaete scute-like homologue 2（ASCL2/Hash2）［J］.Mol Cell Biol，2005，25（15）：6404～6414.
33 Zhu R，Yang Y，Tian Y，et al.ASCL2 knockdown results in tumor growth arrest by miRNA-302b-related inhibition of colon cancer progenitor cells［J］.PLoS One，2012，7（2）：e32170.

[1]	LIU Dandan, LI Jinrong, GAO Hanya, LI Shuyi, PENG Fei, XU Liyina, WANG Xin, HAO Baoshan, MAI Zhanhai, WUSIMAN Adelijiang, ABULA Saifuding, LIU Jianhua, ZHANG Wei. Establishment and Verification of Mice Infection Model of Horse/Donkey Source Streptococcus equi subsp.equi in Xinjiang [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(3): 1342-1351.
[2]	SUN Can, WEI Shuo, YAN Yujing, ZHAO Tianyi, ZHU Dexin, DENG Xingmei, GUO Jia, ZHANG Hui, SUN Zhihua. Bioinformatics Analysis of Brucella BspF Protein and Its Effect on Expression of Inflammatory Factors in Macrophages [J]. China Animal Husbandry & Veterinary Medicine, 2024, 51(5): 1846-1856.
[3]	ZHANG Kang, BASANG Zhuza, CIREN Luobu, DANZENG Luosang, NIMA Cangjue, SILANG Wangmu, PUBU Quzhen, DEJI Yuzhen, SUOLANG Quji. Study on the Epigenetic Mechanism of Hypoxic Adaptation in Plateau Animals [J]. China Animal Husbandry & Veterinary Medicine, 2024, 51(4): 1582-1592.
[4]	YANG Yaqiong, CHENG Hongxing, LIANG Mingxia, LIU Yulan, FU Shulin, ZHANG Jing, CHEN Hongbo, REN Hongyan, GUO Ling, CHAO Zhe. Integrated Validation for DNA Methylation and mRNA Expression of 4 Genes in Brain of Glaessrerlla parasuis-infected Piglets [J]. China Animal Husbandry & Veterinary Medicine, 2024, 51(4): 1651-1659.
[5]	ZHANG Silan, ZHI Feijie, DING Jian, SONG Yinjuan, ZHENG Fuying, FU Qiang, CHU Yuefeng, SHI Huijun, XU Jian. Construction of Brucella Effector Protein BspE Gene Deletion Strain and Its Biological Characteristics [J]. China Animal Husbandry & Veterinary Medicine, 2024, 51(12): 5128-5137.
[6]	XIAO Yangyang, MA Zhongchen, LI Ruirui, CHEN Chuangfu, ZHENG Wei, WANG Yong, WANG Pengyan. Bioinformatic Analysis and Polyclonal Antibody Preparation of Brucella Secreted Protein BspI [J]. China Animal Husbandry & Veterinary Medicine, 2022, 49(8): 3112-3121.
[7]	MAI Zhanhai, LU Yabin, LI Jianlong, ZHAO Lu, SUN Danlan, TONG Panpan, KUANG Ling. Effects of Honeysuckle, Forsythia and Their Drug Pair on Resistance Genes and Virulence Genes of Streptococcus equi subsp. equi [J]. China Animal Husbandry & Veterinary Medicine, 2022, 49(2): 709-717.
[8]	ZHU Yixuan, LIU Shipeng, ZHANG Wei, LI Jinrong, PENG Fei, LIN Na, WANG Yuyan, LIU Dandan. Study on the Optimization of Modified Formula for Inhibiting Streptococcus equi subspecies equi and Its Bacteriostatic Effect in Vivo [J]. China Animal Husbandry & Veterinary Medicine, 2022, 49(10): 4085-4096.
[9]	CHEN Jing, WU Yan, LI Mei, YUE Jun, ZHU Erpeng, WEN Ming, ZHANG Shuangxiang, CHENG Zhentao. Prokaryotic Expression of LPPA Protein of Mycoplasma mycoides subsp. capri and Establishment of Indirect ELISA Method [J]. China Animal Husbandry & Veterinary Medicine, 2022, 49(1): 318-327.
[10]	XU Pan, ZHONG De, MA Zheng, LIU Linyu, WANG Yihui, ZHAO Xuting. Identification of Candidate Genes for Body Weight in Sujiang Pigs Based on Genome-wide Differential Methylation Analysis [J]. China Animal Husbandry & Veterinary Medicine, 2021, 48(8): 2889-2900.
[11]	XUE Qian, LI Guohui, ZHANG Huiyong, YIN Jianmei, ZHOU Chenghao, ZHU Yunfen, XING Weijie, SU Yijun, ZOU Jianmin, HAN Wei. Genome-wide DNA Methylation Analysis of Hypothalamus and Ovary in Langshan Chicken [J]. China Animal Husbandry & Veterinary Medicine, 2021, 48(2): 417-424.
[12]	LI Ruirui, MA Zhongchen, ZHANG Hongyang, WANG Zhen, NUSUFU·Nuersailike, WANG Yong, CHEN Chuangfu. Polyclonal Antibody Preparation and Bioinformatics Analysis of Brucella Secreted Protein BspD [J]. China Animal Husbandry & Veterinary Medicine, 2021, 48(2): 676-684.
[13]	REN Xiaoli, FAN Yuying, HUANGFU Heping, DONG Qing, SHI Dongmei, LIU Yun. Research Progress on Epigenetic Regulation Mechanism in Canine Tumor [J]. China Animal Husbandry & Veterinary Medicine, 2021, 48(11): 4319-4326.
[14]	ZHANG Hui, LI Xiaoyu, DI Ran, LIU Yufang, CHU Mingxing. Analysis on Difference of Transcription and DNA Methylation Level of AANAT Gene in Ovary of Tan Sheep at Different Reproductive Stages [J]. China Animal Husbandry & Veterinary Medicine, 2021, 48(1): 226-234.
[15]	DU Jianwen, YANG Xuemei, WANG Dan, ZHU Hua, HE Junmin, ABLAT Sulayman, TIAN Yuezhen, ZHAO Bingru, XU Xinming, FU Xuefeng, HANIKEZI Tulafu, WU Weiwei, TIAN Kechuan, HUANG Xixia. Analysis of DNA Methylation and mRNA Expression Levels of ITGB2 Gene in Different Fineness of Subo Merino Sheep [J]. China Animal Husbandry & Veterinary Medicine, 2020, 47(4): 1009-1017.

DNA Methylation Status of ASCL2 Gene in Lungs of SCNT Cattle and Naturally Reproduced Cattle

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments