ski-Ⅱ促胃癌sgc7901细胞凋亡的实验研究
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江苏省教育厅自然科学基金(05KJD320234);江苏省卫生厅科研课题基金(Z201016)


ski-ii promoting apoptosis of sgc7901 cells
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    摘要:

    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-Ⅱ 5 μmol/l组-ski-Ⅱ 10 μmol/l组-ddp 2.5 mg/l组-ddp 2.5 mg/l+ski-Ⅱ 5 μmol/l组和ddp 2.5 mg/l+ski-Ⅱ 10 μmol/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-

    Abstract:

    objective:to study the effect and mechanism of sphingosine kinase inhibitor ski-Ⅱ on the apoptosis of human gastric cancer cell sgc7901. methods:sgc7901 cells were cultured and treated with ski-Ⅱ alone or in combination with cisplatin(ddp).the apoptosis of the tumor cells was analyzed by flow cytometry;the morphological changes were observed by electron microscopy;the expression of sphk1,nf-κb,bcl-2 and bax were detected by immunocytochemistry and western blot. pearson analysis was used to detect the correlations between sphk1 and nf-κb,nf-κb and bcl-2. results:after treated for 48 h,the apoptosis rates in sgc7901 treated with ski-Ⅱ 5,10 μmol/l,ddp 2.5 mg/l,ddp 2.5 mg/l+ski-Ⅱ 5 μmol/l and ddp 2.5 mg/l+ski-Ⅱ 10 μmol/l were (40.39±1.06)%,(45.58±0.75)%,(47.27±1.13)%,(53.64±1.11)% and (66.98±2.32)%,respectively,compared with the control group[(18.46±1.64)%],the differences were significant(p < 0.05). apoptotic bodies were found in sgc7901 cells by electron microscopy. the expression of bax was increased,but the expression of sphk1,nf-κb and bcl-2 was decreased. and the correlations between sphk1 and nf-κb,nf-κb and bcl-2 were positive. conclusion:ski-Ⅱ could induce the apoptosis of sgc7901 cells through down-regulating the expression of sphk1 and nf-κb,and increasing the ratio of bax/bcl-2.

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朱祖安,费素娟,刘 磊,孙 旻,张秋月,刘 莹. ski-Ⅱ促胃癌sgc7901细胞凋亡的实验研究[J].南京医科大学学报(自然科学版),2011,(4):463-468

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  • 收稿日期:2011-09-28
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