| 引用本文: | 韩钰,任玉珊,曹春雨,任东明,周永芹,王艳林.用SiRNA沉默SSAT基因表达降低人A549肺癌细胞对抗癌多胺类似物CPENSpm的敏感性[J].中国现代应用药学,2010,27(1):5-10. |
| .Silence of Spermidine/Spermine N1-Acetyltransferase Expression by SiRNA Decreases Sensitivity of Human A549 Lung Cancer Line to Antitumor Polyamine Analogue CPENSpm[J].Chin J Mod Appl Pharm(中国现代应用药学),2010,27(1):5-10. |
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| 摘要: |
| 目的 探讨精脒/精胺乙酰转移酶(Spermidine/Spermine N1-Acetyltransferase，SSAT)基因表达沉默对多胺类似物CPENSpm抗瘤活性的影响。方法 用SiRNA技术获得SSAT基因表达沉默的人A549肺癌细胞株，QT-RT-PCR 法用于分析SSAT基因的表达水平，MTT法检测细胞的存活率，DNA片段化分析和流式细胞/亚凋亡峰分析检测细胞凋亡状况。结果 成功获得SSAT基因表达沉默的人A549肺癌细胞株。用10 μmol·L-1 CPENSpm处理对照细胞24 h可使SSAT mRNA 水平升高23倍，但在SSAT表达沉默的细胞中CPENSpm无此影响。细胞增殖实验发现，SSAT表达沉默的细胞对CPENSpm (0~20 μmol·L-1 )的药物敏感性显著性低于对照细胞。细胞凋亡分析发现，10 μmol·L-1 CPENSpm处理细胞48 h导致对照细胞出现凋亡细胞典型的DNA片段化现象和亚凋亡峰，但在SSAT表达沉默的细胞株中，CPENSpm诱导细胞凋亡的能力明显减弱。结论 CPENSpm诱导肿瘤细胞内SSAT高表达可能是其抗癌药理活性的分子基础之一。 |
| 关键词: 精脒/精胺乙酰转移酶 SiRNA 多胺类似物 肺癌细胞 药物敏感性 |
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| Silence of Spermidine/Spermine N1-Acetyltransferase Expression by SiRNA Decreases Sensitivity of Human A549 Lung Cancer Line to Antitumor Polyamine Analogue CPENSpm |
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HAN Yu REN Yushan CAO Chunyu REN Dongming ZHOU Yongqin WANG Yanlin*
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| Abstract: |
| OBJECTIVE To investigate the effect of expressive silence of spermidine/spermine N1-acetyltransferase (SSAT) on the actitumor activity of polyamine analogue CPENSpm (N1-cyclopropylmethyl-N11-ethylnorspermine). METHODS SiRNA technique was used to silence expression of SSAT gene in human lung cancer line A549. QT-RT-PCR was performed to assay the expression level of SSAT genes. The cell proliferation was detected by MTT assay. The apoptosis of A549 cells were evaluated by DNA fragmentation and Sub-G1/flow cytometry assay. RESULTS The A549 cell line in which SSAT expression was silenced successfully was obtained. Treating A549 control cells by 10 μmol·L-1 CPENSpm for 24 hours resulted in a 23-folds up-regulation of SSAT expression in mRNA level, but no this effect was observed in SSAT-silence cells. MTT assay demonstrated that SSAT-silence decreased the sensitivity of A549 cells to CPENSpm exposure (0-20 μmol·L-1). DNA fragmentation and sub-G1 assay proved the ability of CPENSpm to induce apoptosis only in the control A549 cells but not in the SSAT-silence cell line. CONCLUSION Up-regulation of SSAT by CPENSpm is possibly one of the molecular bases for its antitumor activity. |
| Key words: SSAT SiRNA polyamine analogue lung cancer cell line drug sensitivity |
