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Basic Study for The Gene-Targeted Therapy in Lung cancer

肺癌基因靶向治疗的基础研究

基本信息

批准号：
05670511
负责人：
AKITA Hirotoshi
金额：
$ 1.41万
依托单位：
Hokkaido University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1993
资助国家：
日本
起止时间：
1993 至 1994
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-05670511/
关键词：
Lung Cancer L-myc Antisense DNA Method

项目摘要

We evaluated the antiproliferative effect of L-myc antisense DNA in NCI-H209, a human small cell lung cancer (SCLC) cell line overexpressing the L-myc gene.The synthetic DNA used in the present study was 15mer oligodeoxynucleoside phosphorothioate (OPT), which showed rapid incorporation into NCI-H209 cells, and which localized mainly in the cell nucleus and weakly in the cytoplasm.The exposure of this cell line to s L-myc antisense DNA convering the translational initiation site L-myc proteins inhibited the cell proliferation in a concentration-dependent manner through the concentrations of 1.6muM to 10muM.Furthermore, the growth inhibition by this antisense DNA was correlated with the level of L-myc expression in three different SCLC cell lines including NCI-H209.A mixed sequence control OPT having the same nucleotide length as the antisense OPT but in random, order, did not show any effect on cell proliferation.The sense OPT treatment induced the some inhibition of cell proliferation, which migh be non-sequence-specific inhibition by OPT independent of conventional antisense gene inhibition, which was reported in other systems.Abundant expression of L-myc mRNA and decreased expression of L-myc protein were observed in the antisense-treated NCI-H209 cells, suggesting that the antisense OPT covering the translational initiation site acted by inhibiting ribosomal translation of the target mRNA,rather than by inhibiting transcription from the gene.Together with unique characteristics of the L-myc gene, including 1) a frequently amplified and overexpressed state in SCLC,and 2) very restricted and low level expression in human adult tissues, the present data indicate that L-myc is a good candidate for the target gene for antisense DNA therapy in SCLC based on molecular biological diagonosis.

我们评估了 L-myc 反义 DNA 在 NCI-H209 中的抗增殖作用，NCI-H209 是一种过表达 L-myc 基因的人小细胞肺癌 (SCLC) 细胞系。本研究中使用的合成 DNA 是 15 聚体寡脱氧核苷硫代磷酸酯 (OPT)，它能快速掺入 NCI-H209 细胞中，主要定位于细胞核和细胞核中。将该细胞系暴露于翻译起始位点 L-myc 蛋白的 s L-myc 反义 DNA 中，在 1.6μM 至 10μM 的浓度范围内以浓度依赖性方式抑制细胞增殖。此外，该反义 DNA 的生长抑制与三种不同 SCLC 细胞系中 L-myc 表达水平相关，包括 NCI-H209.与反义OPT具有相同核苷酸长度但随机、顺序的混合序列对照OPT，对细胞增殖没有任何影响。有义OPT处理诱导了对细胞增殖的一定抑制，这可能是OPT的非序列特异性抑制，独立于常规反义基因抑制，这在其他系统中已报道。L-myc mRNA表达丰富，L-myc蛋白表达减少在反义处理的 NCI-H209 细胞中观察到，这表明覆盖翻译起始位点的反义 OPT 通过抑制靶 mRNA 的核糖体翻译而不是抑制基因的转录来起作用。结合 L-myc 基因的独特特征，包括 1) 在 SCLC 中频繁扩增和过表达状态，以及 2) 在人类成人组织中非常有限和低水平的表达，目前数据表明，基于分子生物学诊断，L-myc是SCLC反义DNA治疗的良好候选靶基因。