青蒿琥酯抑制人头颈部鳞状细胞癌增殖及诱导凋亡的机制

摘要： 目的 探讨天然小分子化合物青蒿琥酯 （Akt） 通过诱导人头颈部鳞癌细胞凋亡而抑制肿瘤生长的分子机制。方法 培养人头颈部鳞癌细胞系 UM-SCC-10A， 利用四甲基偶氮唑蓝 （MTT） 法检测 Akt 半数抑制浓度 （IC50 ）；在荧光显微镜下观察不同浓度 （0、 2.5、 5、 10、 20、 40 μmol/L） Akt 对细胞形态的影响； 流式细胞仪检测细胞周期及凋亡情况； 免疫印迹 （Western blot） 分析 Akt 诱导凋亡相关蛋白和细胞周期调节因子表达水平的变化。结果 Akt 对 UM- SCC-10A 细胞的生长有明显抑制作用， 且生长抑制率随药物浓度增加而增加； 当 Akt 处理细胞 48 h 时， IC50 为 15.01 μmol/L。荧光显微镜下， 使用细胞核染料 Hoechst33258 观察到细胞核内出现凋亡小体； 流式细胞仪分析显示 Akt 诱导细胞周期阻滞在 G1 期， 细胞出现大量凋亡； Western blot 结果显示 P53、 P21 蛋白表达量上调、 细胞周期蛋白 D （Cy⁃ cline D） 下调； 线粒体途径诱导的 Bcl-2 相关 X 蛋白 （Bax）、 细胞色素 C （cytochrome C） 及半胱氨酸天冬氨酸蛋白酶-3 （caspase-3） 表达上调， B 细胞淋巴瘤/白血病-2 （Bcl-2）、 procaspase-3 表达下调， 线粒体膜电位降低。结论 Akt 经由线粒体途径诱导 UM-SCC-10A 细胞凋亡， 阻滞细胞周期于 G1期， 进而抑制肿瘤细胞增殖。

The effects and mechanism of artesunate inhibiting the proliferation and inducing the apoptosis of UM-SCC-10A cells

Abstract： Objective To study the influence of artesunate (Akt) on the proliferation and apoptosis of human head and neck squamous cell carcinoma (HNSCC), and to explore its molecular mechanism thereof. Methods The HNSCC cell line,UM-SCC-10A cells, was cultured in vitro. The Inhibitory concentration 50 (IC50) was examined by MTT assay. The cell mor⁃ phological changes were observed under inverted light micro-scope after being interfered by 0, 2.5, 5, 10, 20 and 40 μmol/L Akt. Cell cycle changes and apoptosis were measured by flow cytometry. And the expression of cell cycle regulators and apop⁃ totic associated protein were detected by Western blot assay. Results MTT assay demonstrated that Akt significantly inhib⁃ ited the proliferation of UM-SCC-10A cells in dose-dependent manner. After UM-SCC-10A cells were treated with Akt for 48 h, IC50 was 15.01 μmol/L. Morphological changes of cell apoptosis such as karyopyknosis and conglomeration were ob⁃ served by Hoechst 33258 staining. Flow cytometry showed that the apoptosis was associated with cell cycle arrest during the G1 phase. Western blot analysis showed that p53 and p21 protein was up-regulated and Cyclin D protein was down-regulat⁃ ed. Furthermore, results revealed that Bcl-2 associated X protein induced by a mitochondrial pathway, cytochrome C and caspase-3 were up-regulated, and Bcl-2 and procaspase-3 were down-regulated. The mitochondrial membrane potential was reduced. Conclusion Artesunate can induce apoptosis of UM-SCC-10A cells via a mitochondrial pathway, which was associated with cell cycle arrest in the G1 phase. As a result, artesunate has an obvious inhibitory effects on proliferation of UM-SCC-10A cells