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基因的人类小细胞肺癌（SCLC）细胞系。本研究中使用的合成DNA为15mer寡脱氧核苷磷酸硫酸磷酸盐（OPT），OPT）它显示了快速掺入NCI-H209细胞，并且主要局部位于细胞核中，在细胞核中弱地定位于细胞核中。该细胞系暴露于S l-Myc反义DNA转化转化启动位点L-MYC蛋白抑制细胞增殖的转化。以浓度依赖于1.6MUM到10MUM的浓度。furthermore，这种反义DNA的生长抑制与三种不同的SCLC细胞系中的L-MYC表达水平相关具有与反义OPT相同的核苷酸长度，但在随机的顺序中没有显示出对细胞增殖的任何影响。在其他系统中报道的反义基因抑制。在反义治疗的NCI-H209细胞中观察到了L-MYC mRNA的丰富表达和L-MYC蛋白的表达降低，这表明反义选择了反义覆盖翻译起始位点由作用于翻译起始位点。抑制靶mRNA的核糖体翻译，而不是通过抑制基因的转录。与L-MYC基因的独特特征结合，包括1）SCLC中经常放大且过表达的状态，以及2）在2）中非常有限和低水平表达人类成人组织，目前的数据表明，L-MYC是基于分子生物对角病的SCLC中反义DNA治疗的靶基因的良好候选者。