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目的探讨奥曲肽与磷脂酰肌醇3激酶(PI3′K)通道抑制剂wortmannin联合用药对人胃癌细胞生长的影响及作用机制。方法2007年6月至8月在武汉大学人民医院消化实验室培养人胃癌细胞BGC-823,以单用浓度为40nmol/L的wortmannin,奥曲肽4个稀释度10-5、10-4、10-3及10-2g/L及它们各稀释度与wortmannin(40nmol/L)的混合培养液作用24h后噻唑蓝(MTT)比色实验检测细胞存活率,单用奥曲肽稀释度为10-3g/L和wortmannin以及二者的混和液作用12、24、36、48h检测细胞存活率,于24h检测细胞周期及各组p27基因信使核糖核苷酸(mRNA)和蛋白表达。结果单用奥曲肽组随浓度增加,抑制作用增强,浓度为10-3g/L时,抑制作用明显较对照组增强;单用wortmannin,有较强的肿瘤细胞抑制生长作用,且两者联合用药抑制作用更加明显;单用奥曲肽浓度组随时间的延长其抑制效率逐渐下降,与wortmannin共同作用时,未见细胞明显耐药;通过细胞周期分析,奥曲肽和wortmannin均能抑制肿瘤细胞使其生长停留在G1期,两者联合用药抑制作用更强;在联合用药组中p27蛋白表达明显升高,且随奥曲肽浓度增加表达增加;p27基因mRNA表达在各组无明显差异。结论奥曲肽和wortmannin对胃癌细胞BGC-823均有较强的抑制和杀伤作用,奥曲肽的抑制和毒性作用具有时间和浓度的依赖性,两者联合用药抑制和毒性效果更显著。PI3′K抑制剂wortmannin和奥曲肽均能使肿瘤细胞停止生长在G1期。
Objective To investigate the effects of combination of octreotide and phosphatidylinositol 3 - kinase (PI3’K) channel inhibitor wortmannin on the growth of human gastric cancer cells and its mechanism. METHODS: Human gastric cancer cell line BGC-823 was cultured in the digestive laboratory of Wuhan University People’s Hospital from June 2007 to August 2007, wortmannin (40 nmol / L) 3 and 10-2g / L, and their dilutions were mixed with wortmannin (40nmol / L) for 24 hours. MTT assay was used to test the cell viability. The single octreotide dilution was 10-3g / L And wortmannin as well as the mixture of the two were used to detect the cell viability at 12, 24, 36 and 48 hours. The cell cycle and the messenger RNA (mRNA) and protein expression of p27 gene in each group were detected at 24 hours. Results With octreotide alone, the inhibitory effect was enhanced with the concentration increasing. When the concentration was 10-3g / L, the inhibitory effect was stronger than that of the control group. With wortmannin alone, the stronger tumor cells inhibited the growth and the combination of the two The effect was more obvious. The inhibitory efficiency of octreotide alone group decreased gradually with the time prolonging. No significant cell resistance was found when combined with wortmannin. Both octreotide and wortmannin inhibited the growth of tumor cells G1 phase, the combination of the two drugs inhibition stronger; p27 protein expression was significantly increased in combination group, and with the increase of octreotide expression increased; p27 gene mRNA expression in each group no significant difference. Conclusion Both octreotide and wortmannin can inhibit and kill gastric cancer cells BGC-823. The inhibitory and toxic effects of octreotide are time and concentration-dependent. Both PI3’K inhibitor wortmannin and octreotide stopped tumor cells from growing in the G1 phase.