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目的:研究鞘氨醇激酶(sphingosine kinase,SphK)抑制剂SKI-Ⅱ对人胃癌SGC7901细胞凋亡的影响,并探讨其具体作用机制。方法:常规培养人胃癌SGC7901细胞,SKI-Ⅱ单用或联合顺铂(DDP)干预,流式细胞术检测细胞凋亡率;电镜下观察用药后细胞超微结构的改变;免疫细胞化学、Western blot检测药物作用后细胞中Sphk1、NF-κB、Bcl-2、Bax的表达。Pearson相关分析检测Sphk1与NF-κB、NF-κB与Bcl-2表达的相关性。结果:SKI-Ⅱ单用或联合DDP干预48 h后,SKI-Ⅱ5μmol/L组、SKI-Ⅱ10μmol/L组、DDP 2.5 mg/L组、DDP 2.5 mg/L+SKI-Ⅱ5μmol/L组和DDP 2.5 mg/L+SKI-Ⅱ10μmol/L组细胞凋亡率分别为(40.39±1.06)%、(45.58±0.75)%、(47.27±1.13)%、(53.64±1.11)%和(66.98±2.32)%。与阴性对照组比,差异均有统计学意义(P<0.05);电镜下观察到SGC7901细胞胞浆内出现凋亡小体;SGC7901细胞中Bax阳性表达率增加,Sphk1、NF-κB、Bcl-2阳性表达率减少。Sphk1与NF-κB、NF-κB和Bcl-2的表达呈正相关。结论:SKI-Ⅱ可以通过抑制细胞中Sphk1的表达从而下调NF-κB的表达,并升高Bax/Bcl-2的比例诱导人胃癌SGC7901细胞的凋亡。
Objective: To study the effect of sphingosine kinase inhibitor SKI-Ⅱ on the apoptosis of human gastric cancer cell line SGC7901 and to explore its specific mechanism. Methods: The apoptosis rate of SGC7901 cells, SKI-Ⅱ alone or combined with cisplatin (DDP) were detected by flow cytometry. The changes of ultrastructure were observed under electron microscope. Immunocytochemistry, Western blot blot was used to detect the expression of Sphk1, NF-κB, Bcl-2 and Bax in the cells after drug treatment. Pearson correlation analysis detected the correlation between Sphk1 and NF-κB, NF-κB and Bcl-2 expression. RESULTS: After treated with SKI-Ⅱ alone or in combination with DDP for 48 h, the cells in SKI-Ⅱ5μmol / L group, SKI-Ⅱ10μmol / L group, DDP 2.5 mg / L group, DDP 2.5 mg / L + SKI- The apoptotic rate in the 2.5 mg / L + SKI-Ⅱ 10 μmol / L group was (40.39 ± 1.06)%, (45.58 ± 0.75)%, (47.27 ± 1.13)%, (53.64 ± 1.11)% and (66.98 ± 2.32) %. (P <0.05). The apoptotic bodies were found in the cytoplasm of SGC7901 cells under electron microscope. The positive rate of Bax expression in SGC7901 cells was increased while the expression of Sphk1, NF-κB and Bcl- 2 positive expression rate decreased. Sphk1 was positively correlated with the expression of NF-κB, NF-κB and Bcl-2. Conclusion: SKI-Ⅱ can down-regulate the expression of NF-κB and increase the ratio of Bax / Bcl-2 to induce the apoptosis of human gastric cancer cell line SGC7901 by inhibiting the expression of Sphk1.