高内涵技术筛选抑制GH3细胞的天然黄酮类化合物的研究

Study on Screening Natural Flavonoids Inhibiting GH3 Cells by High-content Screening Technology

目的利用高通量的高内涵筛选(high-content screening,HCS)技术,从天然黄酮类化合物中筛选出对大鼠生长激素腺瘤GH3细胞具有生长抑制作用的化合物并对其作用机制进行初步探讨。方法 HCS技术检测13种天然黄酮类化合物处理GH3细胞24、48 h后细胞的生长形态及增殖活力。不同浓度的黄酮类化合物处理GH3细胞24 h后经Annexin V-FITC/PI双染,流式细胞仪分析化合物诱导细胞凋亡的情况。结果利用HCS技术从13种化合物中筛选出了对GH3细胞生长抑制效果最显著的化合物异硫氰酸烯丙酯,该化合物能够时间依赖性地抑制GH3细胞增殖,并显著呈剂量依赖性地诱导GH3细胞凋亡。结论 HCS技术能直观清晰地观察药物对GH3细胞形态和细胞生长的影响,是一种体外大规模药物筛选的可靠技术方法。筛选出的化合物异硫氰酸烯丙酯可通过诱导细胞凋亡,抑制GH3细胞增殖。

Objective To screen the natural flavonoids inhibiting growth hormone adenoma GH3 cells using the highcontent screening（HCS） technology, and explore their mechanism of action. Methods After treated with 13 natural flavonoids for 24 and 48 h, the growth morphology and proliferation activity of GH3 cells were determined by HCS.Simultaneously, the apoptosis ratios of GH3 induced by natural flavonoids were detected by Annexin V-FITC/PI.Apoptosis of GH3 cells induced by compounds were detected by flow cytometry. Results Allylisothiocyante screened from 13 natural flavonoids could effectively inhibit the proliferation of GH3 cells in a time-dependent manner and induced apoptosis of GH3 cells dose-dependently. Conclusion High-content screening technology is a powerful tool and reliable method for screening drugs via tracking the in vitro antitumor function of drugs in the morphological changes and proliferation of GH3 cells. The compound allylisothiocyante screening by HCS inhibited the proliferation of GH3 cells through inducing cell apoptosis.