Inhibition of glioma growth by tumor-specific activation of double-stranded RNA-dependent protein kinase PKR
Inhibition of glioma growth by tumor-specific activation of double-stranded RNA-dependent protein kinase PKR
复制标题
DOI：
10.1038/nbt730
复制
发表时间：
2002-09-01
影响因子：
46.9
通讯作者：
Levitzki, A
中科院分区：
文献类型：
--
作者：
Shir, A;Levitzki, A
激活的双链RNA （dsRNA）依赖蛋白激酶PKR是一种有效的生长抑制蛋白，主要在病毒感染的细胞中激活，诱导细胞死亡。在这里，我们研究PKR的选择性激活是否可以用来杀死表达含有缺失或染色体易位的突变基因的癌细胞。我们发现，与缺失或易位旁的片段互补的反义RNA （AS）可以产生足够长度的dsRNA分子，以激活PKR并诱导与突变但非野生型mRNA杂交后的细胞死亡。使用表达表皮生长因子受体（EGFR）截断形式Delta(2-7) EGFR的U87MGDeltaEGFR细胞系，我们发现与独特的外显子1至8连接处互补的39核苷酸（nt） AS RNA的表达在体外和体内导致含有截断EGFR的细胞选择性死亡，但不影响表达野生型EGFR的细胞。肿瘤细胞植入后注入表达39-nt AS序列的慢病毒载体，可明显抑制小鼠脑内胶质母细胞瘤的生长。这种pcr介导的杀伤策略可能对治疗许多表达独特RNA物种的癌症有用。
Activated double-stranded RNA (dsRNA)-dependent protein kinase PKR is a potent growth inhibitory protein that is primarily activated in virally infected cells, inducing cell death. Here we investigate whether selective activation of PKR can be used to kill cancer cells that express mutated genes containing deletions or chromosomal translocations. We show that antisense (AS) RNA complementary to fragments flanking the deletion or translocation can produce a dsRNA molecule of sufficient length to activate PKR and induce cell death following hybridization with mutated but not wild-type mRNA. Using the U87MGDeltaEGFR cell line, which expresses a truncated form of epidermal growth factor receptor (EGFR), Delta(2-7) EGFR, we found that expression of a 39-nucleotide (nt) AS RNA complementary to the unique exon 1 to 8 junction caused selective death of cells harboring the truncated EGFR both in vitro and in vivo but did not affect cells expressing wild-type EGFR. A lentiviral vector expressing the 39-nt AS sequence strongly inhibited glioblastoma growth in mouse brain when injected after tumor cell implantation. This PKR-mediated killing strategy may be useful in treating many cancers that express a unique RNA species.