中华皮肤科杂志 ›› 2005, Vol. 38 ›› Issue (12): 751-754.

• 论著 • 上一篇    下一篇

hTERT反义寡核苷酸抑制Hut78细胞端粒酶活性及诱导细胞凋亡的研究

徐秀莲1, 戚金亮2, 尚淑贤1, 陈声利1, 曾学思1, 孙建方1   

  1. 1. 中国医学科学院、中国协和医科大学皮肤病医院 南京 210042;
    2. 南京大学生命科学院
  • 收稿日期:2004-12-15 发布日期:2005-12-15
  • 通讯作者: 孙建方,E-mail:sunjf@hotmail.com E-mail:sunjf@hotmail.com
  • 基金资助:
    国家教委高校博士点基金资助项目(20030023055)

Inhibition of Telomerase Activity and Induction of Cell Apoptosis in Cutaneous T-cell Lymphoma Cell Line by Human Telomerase Reverse Transcriptase Gene Antisense Oligodeoxynucleotide

XU Xiu- lian1, QI Jin-liang2, SHANG Shu-xian1, CHEN Sheng-li1, ZENG Xue-si1, SUN Jian-fang1   

  1. Institute of Dermatology, Chinese Academy of Medical Sciences & Peking Union Medical College, Nanjing 210042, China
  • Received:2004-12-15 Published:2005-12-15

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摘要： 目的 探讨人端粒酶逆转录酶(hTERT)基因反义寡核苷酸对皮肤T细胞淋巴瘤(CTCL)细胞株端粒酶活性及细胞生长的影响,为CTCL基因治疗提供新的基因靶点。方法 采用脂质体介导的基因转染方法,将不同浓度(10μmol/L、20μmol/L、30μmol/L)的寡核苷酸分别导入CTCL细胞株Hut78中;分别于不同的时间,应用端粒酶重复序列扩增及酶联免疫吸附方法(PCR-ELISA)、逆转录-聚合酶链反应(RT-PCR)、流式细胞仪技术等动态观察转染细胞中端粒酶活性、hTERT mRNA表达以及细胞凋亡的变化。结果 经hTERT反义寡核苷酸(AODN)作用72h后,细胞端粒酶的活性明显下降,hTERT mRNA的表达减弱,并可观察到细胞凋亡,凋亡细胞发生率为13.05%。细胞生长抑制作用有明显的时效性,以30μmol/L、72h时下降最明显。而有义寡核苷酸(SODN)组和对照组以上指标均无明显变化(P>0.05)。结论 hTERT反义寡核苷酸可显著抑制CTCL细胞的端粒酶活性,抑制细胞生长增殖,诱导细胞凋亡。

关键词: 淋巴瘤,T细胞,皮肤, 寡核苷酸类,反义, 端粒,末端转移酶

Abstract: Objective To investigate the effect of human telomerase reverse transcriptase (hTERT) gene antisense oligodeoxynucleotide (AODN) on telomerase activity and cell apoptosis in a cutaneous T-cell lymphoma (CTCL) cell line, Hut78. Methods Different concentrations (10 μmol/L, 20 μmol/L, 30 μmol/L) of telomerase antisense oligodeoxynucleotide were introduced into Hut78 cells by lipofectamine-mediated DNA transfection technique. The expressions of hTERT mRNA and telomerase activity were assessed by reverse transcription-polymerase chain reaction and telomeric repeat amplification protocol, respectively. The proliferation and apoptosis of Hut78 cells were detected by flow cytometry. Results After 72 h of incubation, AODN down-regulated the expression of hTERT mRNA, inhibited telomerase activity significantly,and suppressed the viability of Hut78 cells in a time-dependent manner.Cell growth was most clearly suppressed with 30 μmol/L of AODN after 72 h of incubation. The apoptotic rate was 13.05%. Conclusion Telomerase antisense oligodeoxynucleotide could suppress the viability and proliferation of CTCL cell line by inducing apoptosis of these cells.

Key words: Lymphoma, T-cell, cutaneous, Oligonucleotides, antisense, Telomerase