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目的 探讨端粒酶反义寡核苷酸 (PS- ASODN)与顺铂 (DDP)联合应用对原代胃癌细胞凋亡作用的影响。方法 常规组织块培养法进行胃癌细胞原代纯化培养 ,取第 3代对数生长期细胞分六组进行实验。其中四组在培养 2 4小时及 4 8小时分别加入相同剂量的培养液 ,终浓度为 PS- ASODN3μM,N- ASODN3μM,DDP2 .0μg/ ml。作用 2 4小时后分别在 PS- ASODN组及 N- ASODN组加入终浓度为 2 .0 μg/ ml的 DDP;分别于培养后 2 4、4 8、72及 96小时收集各组细胞。以台盼蓝拒染法计算各组细胞生长抑制率 ,观察 PS- ASODN联合 DDP对原代胃癌细胞生长的影响 ;流式细胞学观察细胞凋亡率及细胞周期变化。结果 终浓度为 3μM的 PS- ASODN作用于原代胃癌细胞 2 4小时后加入 DDP 2 .0μg/ m l,能明显抑制胃癌细胞增殖 ,流式细胞学可检测到凋亡峰 ,细胞受阻于G0 / G1 期 ,作用 4 8及 72小时的凋亡细胞百分率 (35 .1%、4 5 .7% )明显高于 N- ASODN组、PS- ASODN组、DDP组及 N- ASODN+DDP组 ,差异有显著性 (P<0 .0 5 )。其作用呈时间依赖性及序列特异性。结论 以端粒酶 RNA模板区为靶点的 PS- ASODN可促进 DDP诱导的胃癌细胞凋亡 ,对胃癌具有治疗价值
Objective To investigate the effect of combined application of telomerase antisense oligonucleotide (PS-ASODN) and cisplatin (DDP) on the apoptosis of primary gastric cancer cells. Methods The primary culture of gastric cancer cells was performed by conventional tissue culture method. The third generation of logarithmic growth phase cells were divided into six groups for experiment. The four groups were cultured in the same culture medium for 24 hours and 48 hours respectively, and the final concentration was PS-ASODN3μM, N-ASODN3μM and DDP2.0μg / ml. After 24 hours of treatment, DDP at a final concentration of 2.0 μg / ml was added to PS-ASODN and N-ASODN groups, and the cells were harvested at 24, 48, 72 and 96 hours after culture. The growth inhibition rate of each group was calculated by trypan blue exclusion method. The effect of PS-ASODN combined with DDP on the growth of primary gastric cancer cells was observed. The apoptosis rate and cell cycle were observed by flow cytometry. RESULTS: PS-ASODN with a final concentration of 3μM could significantly inhibit the proliferation of gastric cancer cells by adding DDP 2.0μg / ml 24 hours after the primary gastric cancer cells were treated with PS-ASODN. The apoptotic peak was detected by flow cytometry and the cells were blocked by G0 / G1 phase, the percentage of apoptotic cells at 48 and 72 hours (35.1%, 45.7%) was significantly higher than that in the group of ASODN, PS-ASODN, DDP and N-ASODN + DDP There was significant (P <0.05). Its role in time-dependent and sequence-specific. Conclusions PS-ASODN targeting telomerase RNA template region can promote the apoptosis of gastric cancer cells induced by DDP and has therapeutic value for gastric cancer