Downregulation of human polo-like kinase activity by antisense oligonucleotides induces growth inhibition in cancer cells
Downregulation of human polo-like kinase activity by antisense oligonucleotides induces growth inhibition in cancer cells
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DOI：
10.1038/sj.onc.1205412
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发表时间：
2002-05-09
期刊：
影响因子：
8
通讯作者：
Strebhardt, K
中科院分区：
文献类型：
--
作者：
Spänkuch-Schmitt, B;Wolf, G;Strebhardt, K
Polo样激酶（PLK）在调节有丝分裂进程的几个阶段中的核心作用已经在几个物种中产生。PLK 1的过表达在迄今为止分析的大多数人类肿瘤中观察到。PLK 1过表达是患有非小细胞肺癌、头颈部肿瘤、食管癌和黑色素瘤的患者的负性预后因子。为了确定PLK 1对人细胞有丝分裂进程和肿瘤细胞生长的作用，测试了硫代磷酸反义寡核苷酸（ASO）以选择性下调MDA-MB-435（乳腺癌）、HeLa S3（宫颈癌）和A549（非小细胞肺癌）细胞中PLK 1的表达。鉴定了以剂量依赖性和序列特异性方式抑制PLK 1 mRNA和蛋白的ASO。这种方法还导致PLK 1丝氨酸/苏氨酸激酶活性降低。癌细胞中PLK 1水平的下调适度改变了细胞周期进程，G（2）/M细胞的百分比升高（20-30%）。此外，PLK 1蛋白减少的细胞获得具有多个中心体的圆形表型。此外，阿索处理导致细胞培养物中的有效抗增殖作用。对A549细胞的体内抗肿瘤活性相当大。这项研究表明，针对PLK 1的反义抑制剂在耐受性良好的剂量可以被认为是一种癌症治疗剂。
A central role for polo-like kinases (PLK) in regulating several stages of mitotic progression has been born out in several species. Overexpression of PLK1 is observed in the majority of hitherto analysed human tumors. PLK1 overexpression is a negative prognostic factor in patients suffering from non-small cell lung cancer, head and neck tumors, esophageal carcinomas and melanomas. In order to define the role of PLK1 for mitotic progression of human cells and for neoplastic cell growth, phosphorothioate antisense oligonucleotides (ASOs) were tested to selectively downregulate PLK1 expression in MDA-MB-435 (breast cancer), HeLa S3 (cervical carcinoma) and A549 (non-small cell lung cancer) cells. ASOs were identified which suppress PLK1 mRNA and protein in a dose-dependent and sequence-specific manner. This approach also led to reduced PLK1 serine/threonine kinase activity. Downregulation of cellular PLK1 levels in cancer cells altered cell cycle progression moderately with an elevated percentage (20-30%) of cells in G(2)/M. Furthermore, cells with reduced PLK1 protein gained a rounded phenotype with multiple centrosomes. Moreover, ASO treatment resulted in potent antiproliferative effects in cell culture. Considerable antitumor activity was observed in vivo against A549 cells. This study suggests that antisense inhibitors targeted against PLK1 at well tolerated doses may be considered as a cancer therapeutic agent.