小鼠NIS蛋白的生物信息学特征及其在乳腺中的表达分析

doi:10.16431/j.cnki.1671-7236.2020.05.003

Abstract

Abstract: In order to explore the structure,function and expression characteristics of Na⁺/I^- symporter (NIS) protein of mammary gland in mouse,NIS gene was amplified from mammary gland in mouse,after sequencing,NIS protein was analyzed by bioinformatics software in this study.The transcription and expression level of NIS gene were detecteded in different stages of mammary gland development by Western blotting,immunohistochemistry and semi-quantitative PCR.The results showed that the CDS sequences of NIS gene was 1 857 bp,and the homology was 80.40% between Homo sapiens and Mus musculus,the conservation was stronger.There were 618 amino acid residues of NIS protein in mouse,with a molecular weight of 65.7 ku,the molecular formula was C₂₉₉₆H₄₇₀₁N₇₄₉O₈₃₃S₃₂,the theoretical isoelectric point was 7.44,the half-life was 30 h,the extinction coefficient was 102 510,the unstability coefficient was 29.31,and the fat solubility coefficient was 105.91.It was rich in hydrophobic amino acids, and was stable hydrophobin.The secondary structure of NIS protein contained alpha helix (41.42%),extended chain (18.28%),beta turn (4.05%) and random coil (36.25%),which was consist with the tertiary structural results.Western blotting results showed that the specific hybridization bands appeared in all four periods,with a size of 65.7 ku in line with expected results.The immunohistochemistry results showed that the expression of NIS protein increased gradually from puberty and pregnancy to lactation,reached the peak in lactation,there was a significant difference (P<0.05),and returned to a similar puberty state in degenerative period.The level of NIS mRNA was consistent with NIS protein.This study provided a reference for further exploring the role of NIS protein in mammary gland development and lactation regulation.

Key words: mouse; NIS protein; mammary gland; bioinformatics; expression

CLC Number:

Q786

XU Xiao, JING Yiming, WANG Quanqi, XU Limin, LIU Yufen. Bioinformatics Characteristics and Expression Analysis of NIS Protein of Mammary Gland in Mouse[J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(5): 1307-1317.

References

[1] WRIGHT E M,LOO D D,HIRAYAMA B A,et al.Surprising versatility of Na⁺-glucose cotransporters:SLC5[J].Physiology,2004,19(6):370-376.
[2] 金佰明,祁艳波,钞虹,等.钠碘转运体基因在不同哺乳期大鼠乳腺组织中的表达[J].中华地方病学杂志,2018,37(6):446-449. JIN B M,QI Y B,CHAO H,et al.Expression of sodium iodide symporter in mammary gland tissues of rats at different lactation periods[J].Chinese Journal of Endemiology,2018,37(6):446-449.(in Chinese)
[3] 石凌峰,胡渊,倪银星.NIS与甲状腺疾病的关系研究进展[J].重庆医学,2018,6:836-840. SHI L F,HU Y,NI Y X.Research progress on the relationship between NIS and thyroid diseases[J].Chongqing Medical Journal,2018,6:836-840.(in Chinese)
[4] 许建林,梁晓燕,南永刚,等.NIS mRNA在分化型甲状腺癌中的表达及其临床意义[J].西安交通大学学报(医学版),2016,2:260-263. XU J L,LIANG X Y,NAN Y G,et,al.Expression of sodium iodide symporter mRNA and its clinical significance in differentiated thyroid carcinoma[J].Journal of Xi'an Jiaotong University (Medical Sciences),2016,2:260-263.(in Chinese)
[5] DAI G,LEVY O,CARRASCO N.Cloning and characterization of the thyroid iodide transporter[J].Nature,1996,379(6564):458-460.
[6] 冀宏,严丽,李清怀,等.碘钠同向转运体在分化型甲状腺癌组织的表达及临床意义[J].中华实验外科杂志,2016,33(4):1085-1087. JI H,YAN L,LI Q H,et al.Expression of sodium/iodide symporter in differentiated thyroid cancer and its clinical significance[J].Chinese Journal of Experimental Surgery,2016,33(4):1085-1087.(in Chinese)
[7] DE L V A,DOHAN O,LEVY O,et al.Molecular analysis of the sodium/iodide symporter:Impact on thyroid and extrathyroid pathophysiology[J].Physiological Reviews,2000,80(3):1083-1105.
[8] DOHAN O,DE L V A,PARODER V,et al.The sodium/iodide symporter (NIS):Characterization regulation and medical significance[J].Endocrine Reviews,2003,24(1):48-77.
[9] SPITZWEG C,JOBA W,HEUFELDER A E.Expression of thyroid-related genes in human thymus[J].Thyroid Official Journal of the American Thyroid Association,1999,9(2):133-141.
[10] CHO J Y,LEVEILLE R,KAO R,et al.Hormonal regulation of radioiodide uptake activity and Na⁺/I^- symporter expression in mammary glands[J].Journal of Clinical Endocrinology & Metabolism,2000,85(8):2936-2943.
[11] 赵丽丽,任春宇,李淑兰.极北鲵消化道6种内分泌细胞的免疫组织化学研究[J].动物学杂志,2018,53(4):96-103. ZHAO L L,REN C Y,LI S L.Immunohistochemical study on six kinds of endocrine cells in the digestive tract of salamandrella keyserlingii[J].Chinese Journal of Zoology,2018,53(4):96-103.(in Chinese)
[12] 黄蕤,余海放,匡安仁.钠/碘同向转运体基因获取及其克隆与测序[J].华西医学,2004,19(4):627-628. HUANG R,YU H F,KUANG A R.Cloning and nucleotide sequencing of the sodium/iodide symporter gene of human thyroid follicular cell[J].West China Medical Journal,2004,19(4):627-628.(in Chinese)
[13] 张颖.大鼠钠/碘同向转运体(rNIS)cDNA克隆和在大肠杆菌的表达[D].天津:天津医科大学,2004. ZHANG Y.cDNA cloning and expression of rat sodium/iodide symporter(rNIS) in E.coli[D].Tianjin:Tianjin Medical University,2004.(in Chinese)
[14] 柏雨生.蛋白序列分析模型在膜蛋白跨膜区预测中的应用[D].杭州:浙江理工大学,2015. BAI Y S.The application of the protein sequence analysis model in the prediction of transmembrane domains[D].Hangzhou:Zhejiang Sci-Tech University,2015.(in Chinese)
[15] 李宝宝,张振兴,黄海峰,等.羊源多杀性巴氏杆菌ompW基因的克隆及生物信息学分析[J].中国畜牧兽医,2019,46(1):28-36. LI B B,ZHANG Z X,HUANG H F,et al.Cloning and bioinformatics analysis of ompW gene from Pasteurella multocida in goat[J].China Animal Husbandry & Veterinary Medicine,2019,46(1):28-36.(in Chinese)
[16] TU W,XU C,LU B,et al.Acute exposure to synthetic pyrethroids causes bioconcentration and disruption of the hypothalamus-pituitary-thyroid axis in zebra fish embryos[J].Science of the Total Environment,2016,542(Pt A):876-885.
[17] ZHANG H,LV S,MU Z,et al.Iodised salt contribution to iodine nutrition status of pregnant and lactating women[J].British Journal of Nutrition,2015,114(1):126-133.
[18] ZIMMERMANN M B.The role of iodine in human growth and development[J].Seminars in Cell & Developmental Biology,2011,22(6):645-652.
[19] PORTULANO C,PARODERBELENITSKY M,CARRASCO N.The Na⁺/I^- symporter (NIS):Mechanism and medical impact[J].Endocrine Reviews,2014,35(1):106-149.
[20] CAILLOU B,TROALEN F,BAUDIN E,et al.Na⁺/I^- symporter distribution in human thyroid tissues:An immunohistochemical study[J].The Journal of Clinical Endocrinology & Metabolism,1998,83(11):4102-4106.
[21] WAPNIR IL,GORIS M,YUDD A,et al.The Na⁺/I^- symporter mediates iodide uptake in breast cancer metastases and can be selectively down-regulated in the thyroid[J].Clinical Cancer Research,2004,10(13):4294-4302.
[22] JIANG H L,DEGRADO T R.[¹⁸F]Tetrafluoroborate ([¹⁸F]TFB) and its analogs for PET imaging of the sodium/iodide symporter[J].Theranostics,2018,8(14):3918-3931.
[23] 贾悦,刘超,袁庆欣,等.乳腺组织中钠/碘转运体基因及蛋白的表达与乳腺疾病发生的关系[J].中国组织工程研究,2004,8(11):2076-2077. JIA Y,LIU C,YUAN Q X,et al.Relationship of sodium-iodide symporter gene and protein expression in breast tissue with the occurrence of breast disease[J].Chinese Journal of Clinical Rehabilitation,2004,8(11):2076-2077.(in Chinese)
[24] TAZEBAY U H,WAPNIR I L,LEVY O,et al.The mammary gland iodide transporter is expressed during lactation and in breast cancer[J].Nature Medicine,2000,6(8):871-878.
[25] PERRON B,RODRIGUEZ A M,LEBLANC G,et al.Cloning of the mouse sodium iodide symporter and its expression in the mammary gland and other tissues[J].Journal of Endocrinology,2001,170(1):185-196.
[26] 王婷婷.碘过量对哺乳期大鼠TRα1、TRβ1、NIS和Dio1的影响[D].天津:天津医科大学,2017. WANG T T.Effect of iodide excess on TRα1,TRβ1,NIS and Dio1 on the lactating rats[D].Tianjin:Tianjin Medical University,2017.(in Chinese)
[27] KOGAI T,KANAMOTO Y,LI A I,et al.Differential regulation of sodium/iodide symporter gene expression by nuclear receptor ligands in MCF-7 breast cancer cells[J].Endocrinology,2005,146(7):3059-3069.
[28] BALTZAR A,SVANBORG C,JAGGI R.Apoptotic cell death in the lactating mammary gland is enhanced by a folding variant of alpha-lactalbumin[J].Cellular & Molecular Life Sciences,2004,61(10):1221-1228.

Bioinformatics Characteristics and Expression Analysis of NIS Protein of Mammary Gland in Mouse

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