姜黄素对CD34^+CD38^-KG1a细胞增殖和凋亡的影响及其与白消安的协同效应

Effects of curcumin on proliferation and apoptosis of CD34^+CD38^-KG1a cells and its synergetic effect with busulfan

目的探讨姜黄素对CD34+CD38-KG1a细胞增殖和凋亡的影响以及姜黄素联合白消安对CD34+CD38-KG1a细胞的协同效应。方法流式细胞术检测细胞CD34、CD38表面抗原的表达情况、姜黄素诱导CD34+CD38-KG1a细胞的凋亡情况及其对CD34+CD38-KG1a细胞周期的影响。MTT法检测姜黄素对CD34+CD38-KG1a的增殖抑制作用及其与白消安联合作用的效应。甲基纤维素克隆形成实验检测细胞克隆形成能力。倒置光学显微镜观察细胞凋亡形态。Western blot检测Bcl-2蛋白表达水平。结果 KG1a细胞株中CD34+CD38-KG1a细胞占(98.2±3.2)%。姜黄素对CD34+CD38-KG1a细胞具有增殖抑制作用,呈时间-剂量依赖性,并能降低CD34+CD38-KG1a细胞的克隆形成能力。姜黄素与白消安相互作用系数(CDI)<1。CD34+CD38-KG1a细胞被姜黄素阻滞于G0/G1期,S期细胞明显减少,并能诱导CD34+CD38-KG1a细胞凋亡。凋亡细胞体积变大,细胞结构不清,胞膜边缘粗糙。姜黄素能下调CD34+CD38-KG1a细胞Bcl-2蛋白表达水平。结论姜黄素通过降低细胞克隆形成能力、阻滞细胞于G0/G1期和诱导细胞凋亡,抑制CD34+CD38-KG1a细胞的增殖,姜黄素与白消安具有协同作用。Bcl-2蛋白表达水平下调可能促使姜黄素诱导CD34+CD38-KG1a细胞凋亡。

Objective To investigate the effects of curcumin on proliferation and apoptosis of CD34＋ CD38 KGla cells and its synergetic effect with busulfan on CD34＋ CD38- KGla cells. Methods The expressions of CD34 and CD38 on the surface of KGla cells and the effect of curcumin on the cell cycle and apoptosis in CD34＋ CD38- KGla cells were detected by flow cytometry. MTT assay was used to analyze curcumin＇s inhibitory effects on proliferation and synergistic effect with busulfan on CD34＋ CD38- KGla. Clone formation rate was measured by methylcellulose colony-formation assay. Morphological changes of apoptotic cells were observed with the inverted optical microscope. The expression of Bcl-2 at the protein level was detected by Western blot. Results The percentage of CD34-CD38 KGla was （98.2±3.2）% in KGla cells. Curcumin could inhibit the proliferation in time- and dose-dependent manners and reduce the colony-formation ability of CD34-CD38 KGla. The coefficient of drug interaction between curcumin and busulfan was less than 1. CD34＋CD38 KGla cells were arrested in the G0/G1 phase by decreasing S phase cells. Meanwhile, curcumin induced the apoptosis of CD34＋ CD38-KGla cells. Apoptotic cells became bigger than normal ones, with unclear cell structure and rough edge of cell membrane. Theexpression of Bcl-2 at the protein level was down-regulated by curcumin. Conclusion Curcumin inhibited the proliferation of CD34-CD38 KGla cells by reducing colony-formation ability, arresting cells in the G0/G1 phase and inducing apoptosis. Besides, there was a synergistic effect between curcumin and busulfan in CD34＋ CD38 KGla cells. The down-regulated expression of Bcl-2 at the protein level may be associated with curcumin-induced apoptosis of CD34- CD38 KGla cells.