VPA、SAHA和DAC对子宫内膜癌细胞系抑制作用的研究

摘要

目的:观察丙戊酸(VPA)、软木酰苯胺氧肟酸(SAHA)和5-氮-2-脱氧胞苷(DAC)对子宫内膜癌细胞RL-952和HEC-1-B的增殖抑制作用,及对细胞周期、凋亡情况和上皮型钙黏素(E-cad)基因表达的影响.方法:以VPA、SAHA和DAC作用于子宫内膜癌细胞,应用噻唑蓝(MTr)比色法、流式细胞仪和透射电镜对细胞增殖、周期和凋亡情况进行观察.应用逆转录-聚合酶链反应(RT-PCR)法观察药物作用后细胞内E-cad mRNA表达的情况.结果:VPA、SAHA和DAC作用HEC-1-B和RL-952细胞,随着药物浓度的增加,增殖抑制率明显增加(6.77％～93.25％和3.24％～89.25％,5.61％ ～97.36％和2.56％～87.85％,8.25％～92.54％和5.56％～93.47％)(P＜0.01)；同一浓度药物随着作用时间的延长(24～96小时)对HEC-1-B和RL-952细胞增殖抑制率明显增加(42.41％～82.74％和36.17％ ～71.05％,43.15％～82.09％和40.38％～85.93％,43.26％～91.44％和37.75％ ～ 85.37％)(P＜0.01).HEC-1-B和RL-952细胞G0/G1期所占比例,经VPA、SAHA、DAC作用24小时后均较对照组增高,差异有高度统计学意义(P＜0.01).应用VPA、SAHA和DAC后细胞凋亡发生率均较对照组增高,差异有高度统计学意义(P＜0.01),出现特征性凋亡形态特征.VPA、SAHA和DAC作用后,RL-952和HEC-1-B细胞E-cad mRNA表达上调,较对照组均增加,差异有统计学意义(P＜0.05).结论:VPA、SAHA和DAC可有效抑制子宫内膜癌RL-952和HEC-1-B细胞增殖,有明显的细胞周期阻滞和诱导凋亡作用,对E-cad基因具有明显上调作用.%Objective: To observe the changes of cell cycle,apoptosis and E-cad mRNA expression in RL-952 and HEC-1-B endometrial carcinoma cell lines treated with VPA , SAHA and DAC. Methods; RL-952 and HEC-1 -B cells were cultured in the presence of VPA, SAHA and DAC. The cellular proliferation inhibitory ratio was evaluated by MTT assay and E-cad mRNA expression was detected by RT-PCR. Flow cytometry (FCM) was used to monitor the cell cycle arrest and apoptosis. Transmission electronic micro-scope(TEM) was applied to observe the apoptosis cellular morphology. Results: With increased concentration and extended culture time, the inhibitory rates of tumor cell growth were notably increased by VPA,SAHA and DAC(6.77% ～ 93.25%/3.24% ～ 89.25%/5. 56% ～ 93.47% and 5. 61% ～ 97. 36%/2. 56% -87.85%/8.25% -92.54%, respectively) ( P＜0.01). The inhibitory rates of tumor cells were 42.41% ～ 82.74%/36.17% -71. 05% ,43.15% - 82. 09%/40. 38% - 85. 93% and 37. 75% ～ 85. 37%/43. 26% ～ 91.44% with the extended culture time(24 -96 h). VPA ,SAHA and DAC can enhance the ratio of G0/G1 in cell cycle( P＜0.01) ,and can induce tumor cell apoptosis( P ＜0.01). The expression of E-cad mRNA was higher in cells cultured in VPA ,SAHA and DAC than that in control group (P＜0.05). Conclusions: VPA, SAHA and DAC can upregulate E-cad mRNA expression, inhibit the growth and induce the apoptosis of endometrial carcinoma cells.