INDUCIBLE PRODUCTION OF C-FOS ANTISENSE RNA INHIBITS 3T3 CELL-PROLIFERATION
INDUCIBLE PRODUCTION OF C-FOS ANTISENSE RNA INHIBITS 3T3 CELL-PROLIFERATION
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DOI：
10.1073/pnas.83.13.4794
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发表时间：
1986-07-01
影响因子：
11.1
通讯作者：
NIENHUIS, AW
中科院分区：
文献类型：
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作者：
HOLT, JT;GOPAL, TV;NIENHUIS, AW
通过基因转移技术在小鼠 3T3 细胞中产生与 c-fos mRNA 互补的反义 RNA。构建的转录单位由类固醇诱导型小鼠乳腺肿瘤病毒(MMTV)启动子、有义(5”至3”)或反义(3”至5”)方向的小鼠或人5” c-fos基因片段以及来自人β-珠蛋白基因的剪接和聚(A)信号组成。载体中包含赋予新霉素抗性的基因，以允许分离稳定的转化体。地塞米松引起杂种 MMTV-fos-球蛋白 RNA 的显着诱导。含有反义c-fos RNA的杂合转录物的诱导减少了DNA转移后的集落形成，并抑制了反义转录单位已整合到其中的细胞的增殖。相反，集落形成和细胞增殖并没有被含有有义方向的c-fos RNA序列的杂合RNA的诱导所抑制。这些结果表明，产生反义RNA来抑制基因表达的策略可能有助于描述原癌基因的功能。 c-fos 基因产物似乎在正常细胞分裂中发挥着必需的作用。
Antisense RNA complementary to c-fos mRNA was produced in mouse 3T3 cells by gene transfer techniques. Transcriptional units were constructed consisting of a steroid-inducible mouse mammary tumor virus (MMTV) promoter, mouse or human 5'' c-fos gene fragments in either the sense (5'' to 3'') or antisense (3'' to 5'') orientation, and splice and poly(A) signals from the human .beta.-globin gene. A gene that confers neomycin resistance was included in the vectors to allow isolation of stable transformants. Dexamethasone caused a marked induction of hybrid MMTV-fos-globin RNA. Induction of the hybrid transcript containing antisense c-fos RNA decreased colony formation following DNA transfer and inhibited the proliferation of cells into which the antisense transcriptional unit had been integrated. In contrast, colony formation and cell proliferation were not inhibited by induction of hybrid RNA containing c-fos RNA sequences in the sense orientation. These results indicate that the strategy of generating antisense RNA to inhibit gene expression may be useful in delineating the function of protooncogenes. The c-fos gene product appears to have a required role in normal cell division.