[1]黄丽娟,陈宁,叶静,等.反义肽核酸对促甲状腺激素受体mRNA表达的影响[J].国际放射医学核医学杂志,2012,36(5):310-312,319.[doi:10.3760/cnla.j.issn.1673-4114.2012.05.011]
　HUANG Li-juan,CHEN Ning,YE Jing,et al.The effects of antisense peptide nucleic acid on the expression of thyroid stimulating hormone receptor mRNA[J].International Journal of Radiation Medicine and Nuclear Medicine,2012,36(5):310-312,319.[doi:10.3760/cnla.j.issn.1673-4114.2012.05.011]
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反义肽核酸对促甲状腺激素受体mRNA表达的影响(/HTML)
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《国际放射医学核医学杂志》[ISSN:1673-4114/CN:12-1381/R]

卷:
36
期数:
2012年第5期
页码:
310-312,319
栏目:
分子生物学
出版日期:
1900-01-01

文章信息/Info

Title:
The effects of antisense peptide nucleic acid on the expression of thyroid stimulating hormone receptor mRNA
作者:
黄丽娟1;  陈宁2;  叶静2;  沈炳玲2;  朱云娟3;  孟涛1;  张志洋1;  朱学良2
1. 天健生物制药(天津)有限公司, 天津 300457;
2. 天津医科大学病理生理教研室, 天津 300070;
3. 天津医科大学寄生虫学教研室, 天津 300070
Author(s):
HUANG Li-juan1;  CHEN Ning2;  YE Jing2;  SHEN Bing-ling2;  ZHU Yun-juan3;  MENG Tao1;  ZHANG Zhi-yang1;  ZHU Xue-liang2
TianJian Biotechnologies(Tianjin) Limited Company, Tionjin 300457, China
关键词:
肽核酸类; 受体; 促甲状腺素; RNA; 信使
Keywords:
Peptide nucleic acids; Receptors; thyrotropin; RNA; messenger
DOI:
10.3760/cnla.j.issn.1673-4114.2012.05.011
摘要:
目的 探讨反义肽核酸(asPNA)对大鼠甲状腺细胞的细胞膜表面促甲状腺激素受体(TSHR)表达的影响。方法 设计两种与TSHR mRNA不同片段互补的asPNA,分别为核定位信号-asPNA 1(NLS-asPNA1)和NLS-asPNA2,另外合成一段非相关的干扰肽核酸(NLS-scrPNA)序列,用荧光显微镜观察NLS-asPNA能否进入细胞,采用噻唑蓝法检测asPNA对细胞是否具有毒性作用,然后用实时定量RT-PCR检测NLS-asPNA对TSHR mRNA表达的影响。结果 荧光显微镜观察发现,asPNA可以进入到细胞中去.噻唑蓝法检测发现asPNA对细胞没有毒性作用.实时定量RT-PCR结果显示,NLS-asPNA1和NLS-asPNA2可以抑制TSHR mRNA表达,表现为随着NLS-asPNA与细胞培养时间的延长,TSHR cDNA的拷贝数逐渐下降,且呈现时间依赖性,而NLS-scrPNA对TSHRmRNA的表达没有明显的影响。结论 NLS-asPNA可以进入到细胞中去,并且能够下调TSHRmRNA的表达.
Abstract:
Objective To study the effects of antisense peptide nucleic acid(asPNA) on thyroid stimulating hormone receptor(TSHR) of Fisher rat thyroid cells membrane.Methods Two kinds of asPNA which could hybridized to the TSHR mRNA named as nuclear localization signal-asPNA1 (NLS-asPNA1) and NLS-asPNA2 were designed,and a control PNA with a scramble sequence named as NLS-scrPNA was synthetized.The cellular uptake of peptide nucleic acids were analyzed by fluorescence microscopy,and the toxic effect of asPNA to Fisher rat thyroid cells was evaluated by MTT assay.The effect of NLS-asPNA on the expression of TSHR mRNA was detected by realtime quantitative RT-PCR.Results Fluorescence microscopy indicated that asPNA were taken up by the cells.The results of MTT assay showed that all the asPNA had no toxic effect on the cells.The results of realtime quantitative RT-PCR showed that NLS-asPNA1 and NLS-asPNA2 could inhibit the expression of TSHR mRNA,the copies of TSHR cDNA decreased gradually along with the prolongation of culture time.But NLS-scrPNA had no significant effect on the expression of TSHR mRNA.Conclusion NLS-asPNA could be taken up by the cells and down-regulate the expression of TSHR mRNA.

参考文献/References:

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备注/Memo

备注/Memo:
收稿日期:2012-06-13。
通讯作者:黄丽娟(Email:tech@tj-bio.com)
更新日期/Last Update: 1900-01-01