Specific inhibition of c-myc protein biosynthesis using an antisense synthetic deoxy-oligonucleotide in human T lymphocytes.
Specific inhibition of c-myc protein biosynthesis using an antisense synthetic deoxy-oligonucleotide in human T lymphocytes.
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DOI：
10.4049/jimmunol.140.7.2431
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发表时间：
1988-04
影响因子：
4.4
通讯作者：
A. Harel-Bellan;D. Ferris;M. Vinocour;J. Holt;W. Farrar
中科院分区：
文献类型：
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作者：
A. Harel-Bellan;D. Ferris;M. Vinocour;J. Holt;W. Farrar
与人 c-myc 基因第二外显子 5' 端互补的 15 聚体脱氧寡核苷酸特异性抑制人 T 细胞中的 C-myc 蛋白表达。寡核苷酸无需任何处理即可渗入细胞，并在 2 小时内达到细胞结合的平台期。 [35S]蛋氨酸脉冲标记蛋白的二维分析表明，该寡核苷酸特异性阻断由 PHA 在人静息外周 T 细胞中诱导的 c-myc 蛋白的从头合成，而不损害其他蛋白的整体合成。 c-myc 蛋白合成的特异性抑制阻止了由 PHA 诱导增殖的静息 T 细胞或由 IL-2 诱导的 IL-2 依赖性 T 细胞进入 S 期，如[3H]胸苷掺入所示。增殖的抑制是特异性的，因为用相应的有义寡核苷酸没有观察到这种抑制，并且通过用过量的有义寡核苷酸预孵育细胞来逆转增殖的抑制。这些结果清楚地支持了 c-myc 蛋白在增殖过程中的作用，并表明可通过与编码外显子互补的合成寡核苷酸来阻断诱导蛋白表达。
C-myc protein expression in human T cells was specifically inhibited by a 15-mer deoxy-oligonucleotide complementary to the 5' end of the human c-myc gene second exon. The oligonucleotide penetrates the cells without any treatment, with a plateau of cell association reached in 2 h. The oligonucleotide specifically blocked the de novo synthesis of c-myc protein, induced by PHA in human resting peripheral T cells, without impairing the overall synthesis of other proteins, as shown by two-dimensional analysis of [35S]methionine pulse-labeled proteins. The specific inhibition of c-myc protein synthesis prevented the entry into S phase of resting T cells, induced to proliferate by PHA, or IL-2-dependent T cells induced by IL-2, as shown by [3H]thymidine incorporation. The inhibition of proliferation was specific since it was not observed with the corresponding sense-oligonucleotide and was reversed by preincubation of the cells with an excess of sense oligonucleotide. These results clearly support a role for c-myc protein in the proliferation process and show that inducible protein expression can be blocked by means of synthetic oligonucleotides complementary to a coding exon.