5氮胞苷与丁酸抑制人肝癌细胞转移的实验研究

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目的 了解 5氮胞苷和丁酸对肝癌细胞转移及肝癌细胞蛋白表达的影响。方法 使用 5氮胞苷与丁酸处理人肝癌细胞株HepG2及Hep3B ,观察处理后肝癌细胞转移、粘附及增殖的改变以及细胞周期抑制蛋白p2 1waf1、p2 7kip1和p5 3表达的改变。结果  5氮胞苷和丁酸联合使用能够完全阻断两种肝癌细胞的转移 ,转移细胞数均为 0 /膜 ;而对照组HepG2和Hep3B转移细胞数分别为(91± 17) /膜和 (117± 74) /膜 ,P <0 0 1;两种药物均可抑制肝癌细胞的粘附及增殖 ;接种后 6d 5氮胞苷与丁酸处理组的HepG2细胞数分别为 (13 2± 0 6 )× 10 5和 (6 1± 0 6 )× 10 5,明显少于对照组(2 1 6± 1 0 )× 10 5;Hep3B细胞数分别为 (13 7± 1 1)× 10 5和 (4 6± 0 7)× 10 5,明显少于对照组的(18 0± 1 6 )× 10 5,P <0 0 1。附壁试验显示接种后 12 0min ,5氮胞苷与丁酸处理组HepG2附壁细胞百分比分别为 5 9 6± 2 6和 41 8± 2 1,明显少于对照组 96 0± 1 4;Hep3B附壁细胞百分比分别为6 7 9± 1 7和 6 7 1± 2 1,明显少于对照组的 91 7± 1 6 ,P <0 0 1。同时还发现两种药物分别处理均可使肝癌细胞p2 1waf1和p2 7kip1蛋白表达增加。此两种蛋白可能在 5氮胞苷和丁酸抑制肝癌细胞转移的过程中起作用。结论 以细胞周期抑 Objective To investigate the effect of 5 azacytidine and butyrate on liver cancer cell metastasis and hepatoma cell protein expression. Methods Human hepatocellular carcinoma cell lines HepG2 and Hep3B were treated with 5-azacytidine and butyrate, and the changes of hepatoma cell metastasis, adhesion and proliferation, and the changes of the expression of the cell cycle arrest proteins p2 1waf1, p2 7kip1 and p53 were observed. Results The combination of 5-azacytidine and butyrate could completely block the metastasis of two hepatocellular carcinoma cells, and the number of metastatic cells was 0/membrane; while the number of HepG2 and Hep3B metastasis cells in the control group was (91±17)/membrane and 117 ± 74)/membrane, P <0 01; Both drugs can inhibit the adhesion and proliferation of hepatocellular carcinoma cells; the number of HepG2 cells treated with azacytidine and butyrate at 6 days after inoculation was (13 2 ± 0). 6) × 10 5 and (6 1 ± 0 6) × 10 5, significantly less than the control group (2 1 6 ± 1 0) × 10 5; Hep3B cell counts were (13 7 ± 1 1) × 10 5 and (4 6 ± 0 7) × 10 5, significantly less than the control group (18 0 ± 1 6) × 10 5, P <0 0 1. The results of the mural test showed that the percentage of HepG2 adherent cells of 5 azacytidine and butyrate was 5 9 6 ± 2 6 and 41 8 ± 2 1, respectively, significantly less than the control group of 96 0 ± 1 4 at 12 min after inoculation; Hep3B The percentage of adherent cells was 6 7 9 ± 1 7 and 6 7 1 ± 2 1, respectively, which was significantly lower than that of the control group (91 7 ± 16), P <0 01. At the same time, it was also found that the two drugs can increase the expression of p2 1waf1 and p2 7kip1 protein in hepatoma cells. These two proteins may play a role in the inhibition of metastasis of hepatoma cells by 5-azacytidine and butyrate. Conclusions Cell cycle inhibition
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