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目的研究他克莫司在体外对肝癌细胞HepG2的细胞增殖、细胞周期及细胞周期蛋白A(cyclin A)表达的生物学影响。方法选择肝癌细胞HepG2进行体外培养,经含有不同浓度的他克莫司(低、中和高浓度组剂量分别为50μg/L、100和500μg/L、1 000和3 000μg/L,对照组0μg/L)的培养基干预后,采用四甲基偶氮唑盐微量酶反应比色法(MTT)及流式细胞技术,分别检测细胞增殖、细胞周期及cyclin A水平。结果①中浓度(500μg/L)和高浓度(1 000和3 000μg/L)的他克莫司对HepG2细胞有增殖抑制作用,且抑制作用随浓度的增加而增加,而低浓度的他克莫司则无抑制作用。②他克莫司对HepG2细胞周期的影响:中浓度(500μg/L)和高浓度(1 000和3 000μg/L)的他克莫司作用时,HepG2细胞停止在G0/G1期,从而对肿瘤细胞的生长有抑制作用,且抑制作用随浓度的增加而增加,而低浓度的他克莫司则无抑制作用。③他克莫司可以降低HepG2细胞中cyclin A的表达,且与浓度相关,他克莫司浓度越高,cyclin A表达越少。结论他克莫司在体外对HepG2增殖有抑制作用,其中cyclin A发挥一定的作用。
Objective To investigate the biological effects of tacrolimus on the proliferation, cell cycle and cyclin A expression of HepG2 cells in vitro. Methods HepG2 cells were selected and cultured in vitro. The cells were treated with different concentrations of tacrolimus (50 μg / L, 100 and 500 μg / L, 1 000 and 3 000 μg / L, 0 μg / L) medium, the cell proliferation, cell cycle and cyclin A levels were detected by MTT assay and flow cytometry, respectively. Results ① Tacrolimus with medium concentration (500μg / L) and high concentration (1000 and 3000μg / L) had inhibitory effect on HepG2 cells and the inhibitory effect increased with the increasing concentration, while the low concentration of tacrolimus Moore is no inhibitory effect. (2) Effects of tacrolimus on HepG2 cell cycle: HepG2 cells stopped in G0 / G1 phase when treated with medium (500μg / L) and high concentrations of Tacrolimus (1 000 and 3 000μg / L) The growth of tumor cells was inhibited, and the inhibitory effect increased with the increase of concentration, but the low concentration of tacrolimus had no inhibitory effect. (3) Tacrolimus can reduce the expression of cyclin A in HepG2 cells, and is related to the concentration. The higher the concentration of tacrolimus, the less the expression of cyclin A. Conclusion Tacrolimus can inhibit the proliferation of HepG2 in vitro, and cyclin A plays a role.