沙利度胺及黄芩素对HEL细胞增殖、凋亡及CRBN蛋白表达的影响

Effects of thalidomide and baicalein on the proliferation, apoptosis and expression of CRBN protein in HEL cells

目的观察沙利度胺、黄芩素单用及联合应用对人红白血病（HEL）细胞的增殖、凋亡及CRBN（eere-blon）蛋白表达水平的影响。方法沙利度胺组、黄芩素组、沙利度胺＋黄芩素组（联合组）作用于HEL细胞48h，用CCK8法检测细胞的增殖抑制率，流式细胞仪双染法检测细胞的凋亡率，Westernblot检测细胞CRBN蛋白的表达水平。结果沙利度胺与黄芩素单用及联合应用均可抑制HEL细胞的增殖，与药物浓度均呈正相关（r=0．991、r=0．989、r=0．993，P〈0．01）；3组药物均可诱导HEL细胞凋亡，与药物浓度均呈正相关（r=0．887、r=0．921、r=0．934，P〈0．01）；且沙利度胺与黄芩素联合应用的增殖抑制作用和促凋亡作用比单用沙利度胺、黄芩素明显增强，差异均有统计学意义（P〈0．05）。沙利度胺处理HEL细胞48h降低CRBN蛋白表达，与药物浓度呈负相关（r=-0．975，P〈0．01）；黄芩素处理HEL细胞48h诱导CRBN蛋白表达，与药物浓度呈正相关（r=0．973，P〈0．01）。结论沙利度胺联合黄芩素呈剂量依赖性抑制HEL细胞增殖并诱导凋亡，比单用沙利度胺、黄芩素的作用明显增强，其机制可能与黄芩素上调CRBN蛋白的表达而增强HEL细胞对沙利度胺的敏感性有关。

Objective To observe the effects of thalidomide, baicalein and both on the proliferation, apoptosis and expression of cereblon （CRBN） protein in human erythroleukemia （HEL） ceils. Methods HEL cells were treated with thalidomide, baicalein or both for 48 h. The proliferation inhibitory rate was measured by CCK8 method. The apoptotic rate was detected by flow cytometry. The level of CRBN protein was measured by Western blot. Results Thalidomide, baicalein and both could inhibit the proliferation of HEL cells, which were positively correlated with drug concentrations （ r = 0. 991, r = 0.989, and r = 0. 993, P 〈 0.01 ）. The combined use of thalidomide and baicalein produced remarkably enhanced effects against proliferation and stronger proapoptotic effects than thalidomide or baicalein alone （P 〈 0. 05 ）. The level of CRBN protein in HEL cells was decreased after treatment with thalidomid for 48 h, which was negatively cor- related with drug concentration （r = -0. 975, P 〈0.01 ）. The level of CRBN protein in HEL cells was decreased after treatment with baicalein for 48 h, which was positively correlated with drug concentration （ r = 0. 973, P 〈 0.01 ）. Con- clusions The combined use of thalidomide and baicalein can inhibit the proliferation but induce the apoptosis of HEL cells in a dose - dependent manner compared with thalidomide or baicalein alane, which may be associated with the role of baica- lein to up - regulate the expression of the CRBN protein and enhance the sensitivity of HEL ceils towards thalidomide.