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背景:国内外多年的研究表明稀土化合物在体内、外均具有明显的抑瘤性能。目的:观察三氯化镧对大鼠肝癌细胞细胞周期素D1和周期素依赖性激酶4蛋白表达的影响。设计:对照观察。单位:吉林大学第一临床医院肾病科。材料:实验于2002-06/2003-07吉林大学基础医学院组织胚胎教研室细胞培养室完成。CBRH-7919细胞按三氯化镧剂量不同分为4组,即空白对照组,0.01,0.1,1.0mmol/L三氯化镧组,每组分别培养1,3,5d,每组细胞各接种6复孔。方法:以体外培养大鼠肝癌细胞株CBRH-7919为研究对象,给予0.01,0.1,1.0mmol/L三氯化镧,培养后1,3,5d观察CBRH-7919细胞生长变化。①运用流式细胞术计算各期细胞所占比例。②四甲基偶氮唑蓝实验测定各孔吸光度值(A值)。③免疫细胞化学检测与G1期调控有关的细胞周期素D1和周期素依赖性激酶4的变化情况。主要观察指标:CBRH-7919细胞在不同剂量和不同培养时间的细胞周期素D1和周期素依赖性激酶4的表达情况。结果:①三氯化镧对CBRH-7919细胞生长的作用:0.1,1.0mmol/L三氯化镧组培养后3,5d吸光度值低于空白对照组(3d分别为1.140±0.070,0.706±0.092,1.461±0.087;5d分别为1.888±0.020,0.625±0.037,2.544±0.032),差异有显著性意义(P<0.05,0.001)。②三氯化镧对CBRH-7919细胞G0/G1期百分率的影响:1.0mmol/L三氯化镧组培养后1,3,5dG0/G1期细胞百分率高于空白对照组[分别为(60.70±0.20)%,(39.49±0.67)%;(61.66±0.97)%,(45.56±1.00)%;(69.92±0.18)%,(49.24±0.27)%],差异有显著性意义(P<0.01,0.001);0.1mmol/L三氯化镧组培养后5dG0/G1期细胞百分率高于空白对照组[分别为(58.88±0.73)%,(49.24±0.27)%],差异有显著性意义(P<0.05)。③三氯化镧对CBRH-7919细胞细胞周期素D1和细胞周期素依赖性激酶4阳性表达的影响:0.1,1.0mmol/L三氯化镧组培养后1d细胞周期素D1和细胞周期素依赖性激酶4表达(A)明显低于空白对照组(分别为562±35,453±22,860±82;705±84,680±28,762±16),差异有显著性意义(P<0.05,0.01,0.001)。结论:三氯化镧可通过下调细胞周期素D1和周期素依赖性激酶4,使肿瘤细胞从G1期进入S期受阻,从而抑制CBRH-7919细胞的生长。
Background: Many years of researches at home and abroad show that rare earth compounds have obvious antitumor activity both in vitro and in vivo. Objective: To observe the effects of lanthanum trichloride on the expression of cyclin D1 and cyclin-dependent kinase 4 protein in rat hepatoma cells. Design: Controlled observation. Unit: First Affiliated Hospital of Jilin University, Department of Nephrology. MATERIALS: The experiment was performed in the cell culture room of Department of Embryology, School of Basic Medicine, Jilin University from June to July 2003. CBRH-7919 cells according to different doses of lanthanum trichloride divided into four groups, namely blank control group, 0.01,0.1,1.0mmol / L lanthanum trichloride group, each group were cultured 1,3,5 d, each group of cells were inoculated 6 complex hole. Methods: The rat hepatoma cell line CBRH-7919 was cultured in vitro. The growth of CBRH-7919 cells was observed at 1, 3, 5 days after culture with 0.01, 0.1 and 1.0 mmol / L lanthanum trichloride. ① using flow cytometry to calculate the proportion of cells in each phase. (2) Tetramethylbenzothiazolyl blue assay the absorbance value of each well (A value). ③ Immunocytochemistry was used to detect the changes of cyclin D1 and cyclin-dependent kinase 4 in G1 phase. MAIN OUTCOME MEASURES: The expression of cyclin D1 and cyclin-dependent kinase 4 in CBRH-7919 cells at different doses and different culture times. Results: (1) The effect of lanthanum trichloride on the growth of CBRH-7919 cells: The absorbance values at 3 and 5 days of 0.1 and 1.0 mmol / L lanthanum trichloride group were lower than those of the blank control group (3d, 1.140 ± 0.070 and 0.706 ± 0.092 , 1.461 ± 0.087; 5d respectively 1.888 ± 0.020,0.625 ± 0.037,2.544 ± 0.032), the difference was significant (P <0.05,0.001). (2) The effect of lanthanum trichloride on the percentage of G0 / G1 phase in CBRH-7919 cells: The percentage of cells in G0 / G1 phase at 1, 3 and 5d after 1.0mmol / L lanthanum chloride treatment was higher than that in the blank control group [(60.70 ± (P <0.01), (39.49 ± 0.67)%, (61.66 ± 0.97)%, (45.56 ± 1.00)%, (69.92 ± 0.18)% and (49.24 ± 0.27)%, respectively 0.001). The percentage of cells in G0 / G1 phase at 5d after culture with 0.1mmol / L lanthanum trichloride was significantly higher than that in the control group [(58.88 ± 0.73)%, (49.24 ± 0.27)%], respectively <0.05). (3) The effect of lanthanum trichloride on the expression of cyclin D1 and cyclin-dependent kinase 4 in CBRH-7919 cells: Cyclin D1 and cyclin D1 The expression of kinases 4 (A) was significantly lower than that of the blank control group (562 ± 35,453 ± 22,860 ± 82 and 705 ± 84,680 ± 28,762 ± 16 respectively), with significant difference (P <0.05,0.01,0.001). CONCLUSION: Lanthanum trichloride can inhibit the growth of CBRH-7919 cells by down-regulating cyclin D1 and cyclin-dependent kinase 4 and blocking tumor cells from G1 phase to S phase.