化学学报 ›› 2014, Vol. 72 ›› Issue (4): 473-480.DOI: 10.6023/A13101092 上一篇    下一篇

研究论文

钌配合物稳定端粒DNA的作用及其诱导细胞凋亡分子机制的研究

刘莹a, 陈小曼a, 张朗棋a, 孙冬冬a, 周艳晖a, 陈兰美a,b, 刘杰a   

  1. a 暨南大学化学系 广州 510632;
    b 广东医学院药学院 湛江 524023
  • 收稿日期:2013-10-25 出版日期:2014-04-14 发布日期:2014-01-14
  • 通讯作者: 刘杰 E-mail:tliuliu@jnu.edu.cn (Jie Liu); Tel./Fax :+86-20-8522-0223 E-mail:tliuliu@jnu.edu.cn
  • 基金资助:

    项目受国家自然科学基金(Nos.21171070,21371075)、广东省科技计划项目(No.c1211220800571)及广东省自然科学基金和中央高校基本科研业务费专项资金资助.

Stabilization of Telomere DNA, and Mechanism of Apoptosis of Tumor Cells Induced by Ruthenium Complexes

Liu Yinga, Chen Xiaomana, Zhang Langqia, Sun Dongdonga, Zhou Yanhuia, Chen Lanmeia,b, Liu Jiea   

  1. a Department of Chemistry, Jinan University, Guangzhou 510632;
    b School of Pharmacy, Guangdong Medical College, Zhanjiang 524023
  • Received:2013-10-25 Online:2014-04-14 Published:2014-01-14
  • Supported by:

    Project supported by the National Natural Science Foundation of China (Nos. 21171070, 21371075), the Planned Item of Science and Technology of Guangdong Province (No. c1211220800571), the Natural Science Foundation of Guangdong Province and the Fundamental Research Funds for the Central Universities.

端粒酶在肿瘤细胞中高表达,已经成为抗肿瘤药物的重要靶点,由于很多肿瘤细胞中富含G4-DNA,通过稳定G-四链体DNA的形成来抑制端粒酶的活性已成为抗癌药物的一个新策略. 本文设计了两种钌配合物,调查了这两种钌配合物稳定G4-DNA的能力,发现配合物2稳定端粒 G4-DNA的能力强于配合物1,配合物2能够诱导端粒 G4-DNA发生构型的转化,而配合物1不能诱导G4-DNA发生构型的转化,这项研究结果证明,钌配合物与G4-DNA的作用能力与配体的平面性有关. 在抗肿瘤活性方面,配合物2表现出更强的抗肿瘤活性,尤其是对HepG2细胞具有较强的抑制作用,推测其是以端粒酶为靶点发挥的抗肿瘤作用. 配合物2能够诱导肿瘤细胞凋亡,能诱导G1期细胞阻滞和DNA碎片的形成(细胞凋亡的特征). 据此推测本论文设计的钌配合物是一个潜在的抗肿瘤药物.

关键词: 端粒酶, G4-DNA, 钌配合物, 抗肿瘤活性

Telomerase is highly expressed in tumor cells, which has become an important target of anticancer drugs. Since many tumor cells were rich in G4-DNA, we investigated the capabilities of these two ruthenium complexes to stabilize G4-DNA. Two ruthenium(Ⅱ) complexes were synthesized and characterized via electrospray ionization-mass spectrometry. Since many tumor cells were rich in G4-DNA, we investigated the capabilities of these two ruthenium complexes to stabilize G4-DNA. The interactions of these compounds with G-quadruplex DNA have been studied by fluorescence spectroscopy and circular dichroism (CD) spectroscopy. The stabilization of quadruplex DNA to complex 2 was better than complex 1, and complex 2 can induce telomeric G-quadruplex to occur conformation transformation, while complex 1 cannot. The results showed that the interaction of ruthenium complexes with G-quadruplex DNA was related with the plane of ligand. A novel visual method has been developed for making a distinction between ct-DNA and HTG21 by our Ru complexes binding hemin to form the hemin-G-quadruplex DNAzyme. The results showed that in the presence of complex 1 or 2, HTG21 can fold into a G-quadruplex, but CT-DNA cannot form the G-quadruplex structure. The anticancer activities of these complexes were evaluated by using the MTT assay. Interestingly, the anti-tumor activity of complex 2 exhibited greater inhibition to HepG2 cells, suggesting the ruthenium complexes were much less toxic towards normal cells, and speculated that it targeted the telomeric G-quadruplex was play a role on anti-tumor effect. To further evaluate the characteristics of the death induced by complexes 1 and 2-treated cells staining with Hoechst is analyzed by fluorescence microscopy. These results indicated that complex 2 revealed antiproliferative activities by apoptosis. We also used PI staining and flow cytometry to assess whether the ruthenium complex 2 affected cell cycle progression in HepG2 cells. Complex 2 is a potential antitumor drugs that can induce cancer cell death by acting on cell cycle arrest in G1 phase and the formation of DNA fragments (apoptosis characteristics).

Key words: telomerase, G-quadruplex DNA, ruthenium(Ⅱ) complexes, anti-tumor activity