[1]赵爱国,吴曙光.沉默CDK7基因对HepG2细胞株的抗增殖作用[J].南方医科大学学报,2005,(01):58-61.
 ZHAO Ai-guo,WU Shu-guang.Effect of CDK7 gene silencing against hepatoblastoma HepG2 cell proliferation[J].Journal of Southern Medical University,2005,(01):58-61.
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沉默CDK7基因对HepG2细胞株的抗增殖作用()
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《南方医科大学学报》[ISSN:1673-4254/CN:44-1627/R]

卷:
期数:
2005年01期
页码:
58-61
栏目:
出版日期:
2005-01-01

文章信息/Info

Title:
Effect of CDK7 gene silencing against hepatoblastoma HepG2 cell proliferation
作者:
赵爱国 吴曙光
南方医科大学药物研究所, 广东广州510515
Author(s):
ZHAO Ai-guo WU Shu-guang
Institute of Pharmaceutical Science, Southern Medical University, Guangzhou 510515, China
关键词:
细胞周期蛋白依赖激酶7肝细胞癌反义寡脱氧核苷酸
Keywords:
cyclin-dependent kinase 7hepatoblastomaantisense oligodeoxynucleotides
分类号:
R916.1;R916.2
摘要:
目的 筛选对体外培养HepG2细胞抗增殖作用最强的反义寡脱氧核苷酸(anti-senseoligodeoxynucleotides,ASODN)序列,探讨经二次优化后该ASODN片段的抗增殖作用。方法 根据RNAStructure3.71计算机软件模拟人CDK7mRNA获得的二级结构确定待选ASODN,经OligoWalk程序分析这些片段与mRNA互补时的热力学参数变化,以此优选待筛ASODN,以体外培养人肝癌HepG2细胞为筛选体系获得抗增殖作用最强的ASODN,对该片段分别依次向互补区上、下游错位1~5个碱基获得10条ASODN,经结构优化后进行二次筛选。结果 初筛结果显示互补于人CDK7mRNA第284~303片段的部分硫代修饰ASODN抗增殖作用最强,为(40.4±12.6)%;二次筛选显示互补于第287~306片段的全磷酸硫代修饰ASODN抗增殖作用最强,抑制率为(68.3±2.6)%,半数抑制浓度为(51.9±8.6)nmol/L。结论 筛选得到的ASODN片段可显著抑制体外培养HepG2细胞的增殖,有可能作为先导化合物开发特异性CDK7抑制剂。
Abstract:
Objective To screen the antisense oligodeoxynucleotides (ASODN) that inhibit cultured hepatoblastoma HepG2 cell proliferation, and evaluate the antiproliferative potency of modified ASODN. Methods ASODN sequences were selected based on the secondary structure of human CDK7 mRNA predicted with RNAStructure 3.71 software. The binding thermodynamics of CDK7 mRNA to ASODN was analyzed by OligoWalk program. The sequences with the strongest effect against cultured HepG2 cell proliferation in vitro were selected, and the fragments complementary to 1-5 bases upstream or downstream to the complementary region were structurally modified and screened. Results The partial phosphorothioate ASODN complementary to 284-303 region of human CDK7 mRNA was the most powerful inhibitor, and the antiproliferative activity reached 40.4±12.6%; in the second round of screening, the antiproliferative activity of the full phosphorothioate ASODN complementary to the 287-306 region of the mRNA on HepG2 cells was 68.3±2.6%, with IC50 of 51.9±8.6 nmol/L. Conclusion Proliferation of HepG2 cells can be significantly inhibited by the screened ASODN, which might be used as a lead compound in the development of specific CDK7 inhibitors.

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备注/Memo

备注/Memo:
收稿日期:2004-3-10。
作者简介:赵爱国(1970- ),男,广州中医药大学博士后,电话:020-61648167
通讯作者:吴曙光,男,博士研究生导师,电话:020-61648167
更新日期/Last Update: 1900-01-01