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靶向丝氨酸/苏氨酸激酶Pim-2的反义寡脱氧核苷酸抑制了DU-145细胞的增殖。

Antisense oligodeoxynucleotides targeting the serine/threonine kinase Pim-2 inhibited proliferation of DU-145 cells.

作者信息

Dai Jin-ming, Zhang Shu-qun, Zhang Wei, Lin Ru-xian, Ji Zong-zheng, Wang Sheng-qi

机构信息

Beijing Institute of Radiation Medicine, Beijing 100850, China.

出版信息

Acta Pharmacol Sin. 2005 Mar;26(3):364-8. doi: 10.1111/j.1745-7254.2005.00050.x.

Abstract

AIM

To investigate the effect of antisense oligodeoxynucleotides (ASODN) targeting Pim-2 on cell proliferation of DU-145 cells.

METHODS

Three ASODN targeting Pim-2 were designed and synthesized. After transfection with ASODN, cell proliferation was analyzed using an MTS [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt] assay. In addition, Pim-2 mRNA, protein levels, and cell cycles were examined.

RESULTS

The ASODN designed and synthesized by our laboratory significantly reduced Pim-2 mRNA level and protein content in DU-145 cells. After transfection with ASODN for 48 h, a marked reduction in cell viability was observed in DU-145 cells in a dose-dependent manner. No remarkable apoptosis occurred in cells treated with ASODN compared with control cells. However, it should be noted that G1 phase arrest was clearly observed in ASODN-treated cells.

CONCLUSION

ASODN targeting Pim-2 resulted in a marked reduction in DU-145 cell proliferation, and induction of G1 phase cell cycle arrest is one of the important mechanisms for ASODN to reduce cell growth. Moreover, antisense inhibition of Pim-2 expression provides a new promising therapy target for prostate cancer.

摘要

目的

研究靶向Pim-2的反义寡脱氧核苷酸(ASODN)对DU-145细胞增殖的影响。

方法

设计并合成三种靶向Pim-2的ASODN。用ASODN转染细胞后,采用MTS[3-(4,5-二甲基噻唑-2-基)-5-(3-羧基甲氧基苯基)-2-(4-磺基苯基)-2H-四唑鎓,内盐]法分析细胞增殖情况。此外,检测Pim-2 mRNA、蛋白水平及细胞周期。

结果

本实验室设计合成的ASODN显著降低了DU-145细胞中Pim-2 mRNA水平和蛋白含量。用ASODN转染48小时后,DU-145细胞的活力呈剂量依赖性显著降低。与对照细胞相比,经ASODN处理的细胞未发生明显凋亡。然而,值得注意的是,在经ASODN处理的细胞中明显观察到G1期阻滞。

结论

靶向Pim-2的ASODN导致DU-145细胞增殖显著降低,诱导G1期细胞周期阻滞是ASODN降低细胞生长的重要机制之一。此外,对Pim-2表达的反义抑制为前列腺癌提供了一个新的有前景的治疗靶点。

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