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[摘要]
目的 探索瑞香狼素 (SC)抗肿瘤机制。方法 以 HL- 6 0和 SGC- 790 1为靶细胞 ,用 MTT比色法测定细胞增殖抑制 ,荧光显微镜观察凋亡细胞的形态学改变 ,DNA电泳和流式细胞仪检测DNA断裂 ,免疫组化检测 bcl- 2蛋白表达。结果 含 SC药物血清处理细胞 48h后 ,HL- 6 0细胞增殖呈剂量依赖性抑制 ,并表现出典型的凋亡细胞形态学改变及 DNA断裂 :即染色体聚集、核固缩、断裂及阶梯状 DNA电泳条带 ,G1 期前细胞从 11.7%增至 5 7.4% ;而 SGC- 790 1细胞 bcl- 2蛋白表达率从 78.3%下降到 32 .9%。结论 SC可诱导肿瘤细胞凋亡 ,降低 bcl- 2蛋白表达。
[Key word]
[Abstract]
Object To explore the antitumor mechanism of Stellera chamaejasme Linn. (SC). Methods SC containing-serum (SCCS) was derived from mice pretreated with different doses of SC. Cultured human leukemia HL-60 and human gastric adenocarcinoma SGC-7901 cells were used. Inhibition of proliferation was measured using MTT assay. Morphological assessment of apoptosis was performed with fluorescence microscope. DNA fragmentation was assessed by agarose gel electrophoresis and flow cytometry. Expression of bcl-2 protein was measured with immunohistochemistry. Results Exposure of exponentially growing HL-60 cells to mice serum containing 10% SC (pretreated with SC 3, 6, and 12 g/kg) for 48 h resulted in growth inhibition in a dose-dependent manner. Typical morphological changes of apoptosis and DNA fragmentation in HL-60 cells were induced. "Apobodies" in the apoptotic cells were observed, "ladder" pattern of agarose gel electrophoresis of DNA from these cells was revealed, and the percentage of apoptotic cells with fractional DNA content increased from 11.7% to 57.4%. Treatment with SC containing serum decreased the percentage of SGC-7901 cells of bcl-2 protein positive expression from 78.3% to 32.9%. Conclusion SC could induce apoptosis of HL-60 cells and decrease the expression of bcl-2 protein of gastric adenocarcinoma SGC-7901 cells.
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[基金项目]
国家自然科学基金资助项目No29972054