目的 考察小白菊內(nèi)酯對(duì)于人腦惡性膠質(zhì)瘤細(xì)胞U-87 MG細(xì)胞系的增殖、細(xì)胞周期及凋亡的影響,并探討其作用機(jī)制。方法 采用CCK-8比色法和克隆形成檢測(cè)法檢測(cè)小白菊內(nèi)酯對(duì)U-87 MG的增殖抑制作用;采用碘化丙啶(PI)染色后的流式細(xì)胞儀檢測(cè)小白菊內(nèi)酯處理前后對(duì)細(xì)胞周期、凋亡的影響;采用qPCR以及Western blotting法研究藥物處理后U-87 MG細(xì)胞的Caspase 3、Bax、Cyclin D1的表達(dá)變化以及P53-Ser³⁹²蛋白質(zhì)磷酸化激活。結(jié)果 小白菊內(nèi)酯可明顯抑制U-87 MG的細(xì)胞增殖、克隆形成數(shù)量及克隆大小。處理48 h后,U-87 MG細(xì)胞的S期、G2/M期受到了阻滯,G0/G1期細(xì)胞比例明顯減少,同時(shí)細(xì)胞的凋亡也明顯增多。Caspase 3、Bax基因的mRNA及蛋白表達(dá)明顯上升,而Cyclin D1基因的mRNA表達(dá)明顯下降。同時(shí)P53-Ser³⁹²殘基的磷酸化水平也明顯上升。結(jié)論 小白菊內(nèi)酯可通過(guò)調(diào)節(jié)P53基因的Ser³⁹²殘基磷酸化抑制其下游的細(xì)胞凋亡、細(xì)胞周期途徑上Caspase 3、Bax、Cyclin D1的基因表達(dá),影響細(xì)胞周期及凋亡,從而抑制腫瘤細(xì)胞的增殖。

Objective To study the effects of parthenolide on human malignant glioma cells U-87 MG cell growth, cell cycle regulation, and apoptosis, and explore its mechanism. Methods The inhibition of proliferation on U-87 MG cell of parthenolide were determined using CCK8 and colony formation assay. Effects on cell cycle and apoptosis changes treated by parthenolide were detected by flow cytometer analysis after PI staining. The expressions of cell cycle and apoptosis related proteins, Caspase 3, Bax, Cyclin D1, and P53-Ser³⁹² were analyzed by qPCR and western blotting. Results Parthenolide significantly inhibited cell proliferation, and induced decrease in the colony formation ability inU-87 MG cells. Treated by parthenolide after 48 h, U-87 MG cell were arrested at S and G2/M phases, the cells proportion at G0/G1 phase decreased, and the apoptotic cells increased. Furthermore, parthenolide could increase the mRNA expression of caspase 3, Bax, and P53-Ser³⁹², while decrease the mRNA expression of Cyclin D1. Conclusion Parthenolide can regulate the Ser³⁹² residue phosphorylation of P53 gene, inhibit apoptosis of downstream cells, and expression of caspase 3, Bax, and Cyclin D1 genes, which effects cell cycle and apoptosis, and induce cellular senescence and apoptosis.

廣西壯族自治區(qū)衛(wèi)生廳自籌經(jīng)費(fèi)科研課題(Z2013624)