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目的探讨黄癸素(α-羟基-δ-癸酰乙基磺酸小檗碱)对人乳腺癌MDA-MB-231细胞增殖抑制作用及对核转录因子(NF)-κβ、P65、P53蛋白表达的影响。方法体外培养人乳腺癌MDA-MB-231细胞,分别加入不同终浓度黄癸素(0.4 mg/L、0.8 mg/L、1.6 mg/L、3.2 mg/L、6.4 mg/L)处理作为实验组,未用黄癸素处理的细胞作为对照组。采用噻唑蓝(MTT)比色法、4’,6-二脒基-2-苯基吲哚(DAPI)染色法、碘化丙啶(PI)单染流式细胞术、Annexin VFITC/PI双染流式细胞术、Western bolt法,分别检测黄癸素对乳腺癌MDA-MB-231细胞增殖抑制作用、细胞凋亡形态、细胞周期各时相细胞比例、细胞凋亡水平和NF-κβ、P65、P53蛋白表达水平的影响。结果经黄癸素处理后,5个浓度实验组MDA-MB-231细胞增殖抑制率随黄癸素浓度的增加而递增(P<0.01),呈药物剂量依赖性;不同浓度实验组培养48 h的细胞增殖抑制率明显高于培养24 h(P均<0.01);培养24 h的半抑制浓度(IC50)=3.03 mg/L,培养48 h的IC50=2.00 mg/L。1.6 mg/L和3.2 mg/L两个实验组作用48 h后,随黄癸素浓度的增加(0→1.6 mg/L→3.2 mg/L),MDA-MB-231细胞G0/G1期百分数递增(P<0.01),G2/M期百分数基本不变(P>0.05),S期细胞百分数则递减(P<0.01);细胞凋亡率递增(P<0.01);NF-κβ表达水平递减,P53、P65蛋白表达水平递升;均呈剂量依赖性。结论黄癸素具有明显抑制乳腺癌MDA-MB-231细胞增殖的作用,且呈药物-剂量依赖性,其可能通过下调NF-κβ,上调P65、P53蛋白表达诱导细胞凋亡。
OBJECTIVE To investigate the inhibitory effect of huang decoction on the proliferation of human breast cancer MDA-MB-231 cells and the expression of nuclear factor-kappa B (NF-κB), P65 and P53 influences. Methods Human breast cancer MDA-MB-231 cells were cultured in vitro and treated with different concentrations of flavonoids (0.4 mg / L, 0.8 mg / L, 1.6 mg / L, 3.2 mg / L, 6.4 mg / L) Cells that were not treated with the yellow decanoic acid served as controls. The cells were stained with MTT assay, 4 ’, 6-diamidino-2-phenylindole (DAPI) and propidium iodide (PI) Flow cytometry and Western blot were used to detect the inhibitory effects of CDF on the proliferation of breast cancer MDA-MB-231 cells, the morphological changes of apoptotic cells, the proportion of cells in each phase of the cell cycle, the levels of apoptosis and the expressions of NF-κβ, P65, P53 protein expression levels. Results The proliferation inhibition rate of MDA-MB-231 cells in five concentration groups was increased with the increase of the concentration of theuridine (P <0.01), and the dose-dependent manner. The proliferation of the cells in different concentration groups for 48 h (IC50) = 3.03 mg / L at 24 h and IC50 = 2.00 mg / L at 48 h after culture for 24 h (all P <0.01). The percentage of G0 / G1 phase of MDA-MB-231 cells increased gradually with the increase of the concentration of Huangui Suai (0 → 1.6 mg / L → 3.2 mg / L) after treated with 1.6 mg / L and 3.2 mg / (P <0.01). The percentage of G2 / M phase remained unchanged (P> 0.05), the percentage of S phase cells decreased (P <0.01), the cell apoptosis rate increased (P <0.01) , P65 protein expression levels increased; all in a dose-dependent manner. Conclusion Huang decoction can significantly inhibit the proliferation of breast cancer MDA-MB-231 cells in a dose-dependent manner. It may induce apoptosis by down-regulating NF-κβ, up-regulating P65 and P53 protein expression.