芹菜素对人急性髓性白血病细胞化疗敏感性的增强作用

Chemosensitization effects of apigenin on cisplatin in human acute myeloid leukaemia HL-60 cells

目的观察芹菜素(API)能否增强人急性髓性白血病(HL-60)细胞系细胞对顺铂(DDP)的化疗敏感性。方法取对数生长期的HL-60细胞,分为API组、DDP组、API+DDP组与空白对照组。API组加入API,终浓度为1.0μmol·L-1,DDP组加入DDP,终浓度分别为1.0mg·L-1,2.0mg·L-1,4.0mg·L-1,API+DDP组API终浓度为1.0μmol·L-1,DDP分别为1.0mg·L-1,2.0mg·L-1和4.0mg·L-1,空白对照组仅加入等量完全培养基。采用MTT比色法测定细胞生长抑制率。另取对数生长期的HL-60细胞,分组同上,采用流式细胞术测定细胞凋亡率。另取对数生长期的HL-60细胞,分为3组,前2组加入1.0μmol·L-1API,分别培养24h,48h,对照组仅加入等量完全培养基,采用间接免疫荧光标记流式细胞术观察API对HL-60细胞NF-κB和Bcl-2表达的影响。结果1.0μmol·L-1API无细胞毒作用,不能有效诱导HL-60细胞的凋亡,但它与不同浓度的DDP合用,可增强DDP对HL-60细胞增殖的抑制作用,并使细胞凋亡率明显增加。1.0μmol·L-1API可抑制HL-60细胞NF-κB和Bcl-2的表达(P均<0.01)。结论API可增强DDP对HL-60细胞增殖的抑制作用和诱导凋亡作用,其机制可能与下调NF-κB和Bcl-2的表达有关。

Aim ： To observe the chemosensitization effects of apigenin （API） on cisplatin （DDP） in human acute myeloid leukaemia HL-60 cells. Methods： Ⅰ ： HL-60 cells were allocated into API group, DDP groups, API ＋ DDP groups, and control group. API group was given 1.0 μmg·L^-1 API, DDP groups were given 1.0 mg·L^-1 DDP, 2.0 mg · L^-1 DDP,4.0 mg · L^-1 DDP,respectively, and API ＋ DDP groups were given 1.0 μmol · L^-1 API plus 1.0 mg · L^-1 DDP,2.0 mg· L^-1 DDP,4.0 mg · L^-1 DDP, respectively. Control group was only given culture fluid. The inhibition rate of cell growth was detected using MTT assay, Ⅱ - HL-60 cells were allocated as above method and the apoptosis rate was determined using flow cytometry. Ⅲ ： HL-60 cells were allocated into API groups and control group. API groups were given 1.0 μmol·L^-1 API and cultured for 24 h, 48 h, respectively, and the control group was only given culture fluid. The expressions of NF-κB and Bcl-2 in HL-60 cells were detected using immunofluorescence flow cytometry. Results： 1.0 μmol · L^-1 API had no cytotoxic effects on HL-60 cells and did not induce cell apoptosis but after being combined with DDP, the inhibition rate and the apoptosis rate increased, and the expressions of NF-κB and Bcl-2 were downregulated compared with DDP groups. Conclusion： API promotes proliferative inhibition and apoptotic induction of HL-60 cells by DDP, which may associate with its downregulating the expressions of NF-κB and Bcl-2 protein.