Liu Xing-An
Department of Radiotherapy, People's Hospital of Zhengzhou, Zhengzhou 450003, Henan Province, China.
Int J Ophthalmol. 2012;5(3):272-6. doi: 10.3980/j.issn.2222-3959.2012.03.04. Epub 2012 Jun 18.
To observe the effects of salvianolic acid B (SalB) on in vitro growth inhibition and apoptosis induction of retinoblastoma HXO-RB44 cells.
The effects of SalB on the HXO-RB44 cells proliferation in vitro were observed by MTT colorimetric method. The morphological changes of apoptosis before and after the treatment of SalB were observed by Hoechst 33258 fluorescent staining method. Apoptosis rate and cell cycle changes of HXO-RB44 cells were detected by flow cytometer at 48 hours after treated by SalB. The expression changes of Caspase-3 protein in HXO-RB44 cells were detected by Western Blot.
SalB significantly inhibited the growth of HXO-RB44 cells, while the inhibition was in a concentration-and time-dependent manner. The results of fluorescent staining method indicated that HXO-RB44 cells showed significant phenomenon of apoptosis including karyorrhexis, fragmentation and the formation of apoptotic bodies, etc. after 24, 48 and 72 hours co-culturing of SalB and HXO-RB44 cells. The results of flow cytometer showed that the apoptosis rate and the proportion of cells in S phase were gradually increased at 48 hours and 72 hours after treated by different concentrations of SalB. Western Blot strip showed that the expression of Caspase-3 protein in HXO-RB44 cells was gradually increased with the increase of the concentration of SalB.
SalB can significantly affect on HXO-RB44 cells growth inhibition and apoptosis induction which may be achieved through the up-regulation of Caspase-3 expression and the induction of cell cycle arrest.
观察丹酚酸B(SalB)对视网膜母细胞瘤HXO-RB44细胞体外生长抑制及凋亡诱导作用。
采用MTT比色法观察SalB对HXO-RB44细胞体外增殖的影响。采用Hoechst 33258荧光染色法观察SalB处理前后细胞凋亡的形态学变化。SalB处理48小时后,采用流式细胞仪检测HXO-RB44细胞凋亡率及细胞周期变化。采用Western Blot检测HXO-RB44细胞中Caspase-3蛋白表达变化。
SalB显著抑制HXO-RB44细胞生长,且抑制作用呈浓度和时间依赖性。荧光染色结果显示,SalB与HXO-RB44细胞共培养24、48和72小时后,HXO-RB44细胞出现明显凋亡现象,包括核固缩、核碎裂及凋亡小体形成等。流式细胞仪检测结果显示,不同浓度SalB处理48小时和72小时后,细胞凋亡率及S期细胞比例逐渐升高。Western Blot条带显示,HXO-RB44细胞中Caspase-3蛋白表达随SalB浓度增加而逐渐升高。
SalB可显著影响HXO-RB44细胞的生长抑制和凋亡诱导,其机制可能是通过上调Caspase-3表达及诱导细胞周期阻滞实现的。