Screening and activity evaluation for potential inhibitors of human PLK1

doi:10.3969/j.issn.1006-7795.2014.03.015

Abstract

Abstract:

Objective To find potential small-molecular PLK1 inhibitors through high throughput screening using budding yeast assay and examine the in vitro anti-tumor effects of these compounds. Methods The influences of the positive compounds on cells' proliferation were measured by MTT assays. ELISA assay was employed to explore their inhibition activities on purified recombinant human PLK1. Flow cytometry(FCM) was conducted to determine their effects on cell cycle and apoptosis in Hela cells. The levels of Cyclin B1 and PLK1 were detected by Western blotting. Results We got 3 hits from preliminary screening by yeast assay. In vitro antitumor results revealed that compounds 1 and 3 showed the enhanced inhibition on the cell proliferation of human cancer cell lines with concentration increasing, while the activity of compound 2 was very weak. Compound 1 had lower nanomolar activity against the PLK1 enzyme without influencing its protein expression compared with compound 2 and 3. FCM showed that cells treated with compound 1 were arrested in G2/M phase. Western blotting showed an increase in expression of Cyclin B1 in Hela cells after treated with compound 1. Conclusion The inhibition effect of compound 1 on PLK1 was associated with kinase activity without influencing the PLK1 protein expression. Compound 1 is expected to become one of the anti-cancer drug candidates targeted at PLK1.

Key words: Polo-like kinase 1 inhibitor, anti-cancer, cell cycle, apoptosis, Western blotting

CLC Number:

Zhang Jing, Liu Wei, Chen Yunyu, Si Shuyi. Screening and activity evaluation for potential inhibitors of human PLK1[J]. Journal of Capital Medical University, 2014, 35(3): 337-343.

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URL: https://journal03.magtech.org.cn/Jweb_sdykdxxb/EN/10.3969/j.issn.1006-7795.2014.03.015

https://journal03.magtech.org.cn/Jweb_sdykdxxb/EN/Y2014/V35/I3/337

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