Effect of 2'-O-methyl antisense ORNs on expression of thymidylate synthase in human colon cancer RKO cells.
Effect of 2'-O-methyl antisense ORNs on expression of thymidylate synthase in human colon cancer RKO cells.
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2-O-甲基反义 ORN 对人结肠癌 RKO 细胞中胸苷酸合酶表达的影响。
DOI：
10.1093/nar/29.2.415
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发表时间：
2001
影响因子：
14.9
通讯作者：
Chu,E
中科院分区：
文献类型：
--
作者：
Schmitz,JC;Yu,D;Agrawal,S;Chu,E
胸苷酸合酶 (TS) mRNA 的翻译由其自身的蛋白质终产物 TS 以负向自动调节方式控制。这种调节的破坏会导致 TS 合成增加，并可能导致细胞对 TS 导向的抗癌药物产生耐药性。作为抑制 TS 表达的策略，反义 2'-O-甲基 RNA 寡核糖核苷酸 (ORN) 被设计为直接靶向 TS mRNA 的 5' 上游顺式作用调节元件（核苷酸 80-109）。 30 nt ORN HYB0432 以剂量依赖性方式抑制人结肠癌 RKO 细胞中的 TS 表达，但对 β-肌动蛋白、α-微管蛋白或拓扑异构酶 I 的表达没有影响。TS 表达不受对照正义或错配 ORN 处理的影响。 HYB0504 是一种针对相同核心序列的 18 nt ORN，也抑制 TS 蛋白的表达。然而，寡核苷酸尺寸的进一步减小导致反义活性的丧失。 Following HYB0432 treatment, TS protein levels were reduced by 60% within 6 h and were maximally reduced by 24 h. p53蛋白的表达与TS的表达呈负相关，表明p53的表达可能与细胞内TS的水平直接相关。 Northern 印迹分析表明 TS mRNA 不受 HYB0432 处理的影响。反义处理后 TS 蛋白的半衰期没有变化，表明反义 ORN 的作用机制是通过翻译停滞过程介导的。这些发现表明，针对 TS mRNA 上关键顺式作用元件的反义 ORN 可以特异性抑制 RKO 细胞中 TS 蛋白的表达。
Translation of thymidylate synthase (TS) mRNA is controlled by its own protein end-product TS in a negative autoregulatory manner. Disruption of this regulation results in increased synthesis of TS and may lead to the development of cellular drug resistance to TS-directed anticancer agents. As a strategy to inhibit TS expression, antisense 2′-O-methyl RNA oligoribonucleotides (ORNs) were designed to directly target the 5′ upstreamcis-acting regulatory element (nucleotides 80–109) of TS mRNA. A 30 nt ORN, HYB0432, inhibited TS expression in human colon cancer RKO cells in a dose-dependent manner but had no effect on the expression of β-actin, α-tubulin or topoisomerase I. TS expression was unaffected by treatment with control sense or mismatched ORNs. HYB0504, an 18 nt ORN targeting the same core sequence, also repressed expression of TS protein. However, further reduction in oligo size resulted in loss of antisense activity. Following HYB0432 treatment, TS protein levels were reduced by 60% within 6 h and were maximally reduced by 24 h. Expression of p53 protein was inversely related to that of TS, suggesting that p53 expression may be directly linked to intracellular levels of TS. Northern blot analysis demonstrated that TS mRNA was unaffected by HYB0432 treatment. The half-life of TS protein was unchanged after antisense treatment suggesting that the mechanism of action of antisense ORNs is mediated through a process of translational arrest. These findings demonstrate that an antisense ORN targeted at a criticalcis-acting element on TS mRNA can specifically inhibit expression of TS protein in RKO cells.
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影响因子：
11.2
作者：
P. Johnston;H. Lenz;C. Leichman;K. Danenberg;C. Allegra;P. Danenberg;L. Leichman
通讯作者：
P. Johnston;H. Lenz;C. Leichman;K. Danenberg;C. Allegra;P. Danenberg;L. Leichman
使用通过聚乙二醇与叶酸缀合的脂质体，将针对人表皮生长因子受体的反义寡脱氧核糖核苷酸递送至培养的 KB 细胞中。
DOI：
10.1073/pnas.92.8.3318
发表时间：
1995
影响因子：
11.1
作者：
Susan H. H. Wang;R. J. Lee;G. Cauchon;D. Gorenstein;P. Low
通讯作者：
P. Low
DOI：
10.1089/oli.1.1997.7.151
发表时间：
1997-06-01
期刊：
ANTISENSE & NUCLEIC ACID DRUG DEVELOPMENT
影响因子：
--
作者：
Stein, D;Foster, E;Summerton, J
通讯作者：
Summerton, J
影响因子：
56.9
作者：
SHOICHET, BK;STROUD, RM;PERRY, KM
通讯作者：
PERRY, KM
DOI：
10.1016/s0021-9258(18)82448-6
发表时间：
1993-07
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
K. Keyomarsi;Jonathan M. Samet;Gyongyi Molnar;A. Pardee
通讯作者：
K. Keyomarsi;Jonathan M. Samet;Gyongyi Molnar;A. Pardee