【摘要】 目的 :观察肝素对叉孢素介导的人鼻咽癌细胞的增殖与凋亡的影响 ,以及探讨其可能的分子机制。方法 :应用了MTT法、荧光染色、TUNEL、流式细胞术、琼脂糖凝胶电泳、RT -PCR及Westernblot等方法。结果 :肝素与叉孢素联合应用后对鼻咽癌细胞的增殖及凋亡具有显著的影响。低剂量的肝素与叉孢素联合应用 :细胞生长抑制率显著高于叉孢素单独应用 ;细胞核浓缩 ,核碎裂等形态学变化更加明显 ;TUNEL阳性细胞明显增多 ;凋亡率增加 ;DNA“梯形”变化也清晰可见 ;c-myc ,bax基因的表达明显高于叉孢素单独应用 ,而bcl- 2在整个用药过程中没有改变。结论 :肝素与叉孢素对人鼻咽癌细胞在抗增殖及促凋亡方面有协同作用 ,这可能与c -myc ,bax的表达上调有关 .更多还原

【Abstract】 AIM: To determine whether heparin combined with staurosporine was more effective than either agent alone on anti-proliferation and apoptosis in CNE2 cells, to improve the understanding of the mechanisms by which staurosporine and heparin influence cell proliferation and apoptosis, and to further examine the role of different apoptosis-related oncogenes in regulation of staurosporine and/or heparin -mediated cell proliferation and apoptosis in CNE2 cells. METHODS: Cell viability was determinted by methl-terazolium (MTT). Apoptosis was evaluated by Hoechst 33258 staining, TUNEL, and agarose gel electrophoresis as well as flow cytometry. The expression of bcl-2, bax, c-myc was examined by RT-PCR and Western blot. RESULTS: Staurosporine elicited typical apoptotic morphologic changes. Compared with staurosporine alone, staurosporine plus heparin obviously enhanced the number of TUNEL-positive cells,and the percentage of apoptosis was 2 6%±1 2%, 14 3%±1 5%, 33 2%±0 2%, 57 9%±0 3%, respectively. SubG 1 peaks were detected on flow cytometry, the apoptotic index was enhanced from 3 8% to 56 8%. The DNA laddering was more clearly observed by the treatment with staurosporine plus heparin. Similarly, the expression of bax protein and mRNA, and the level of c-myc protein were overexpressed by the treatment with staurosporine plus heparin, while no significant changes were found in the expression of bcl-2 protein and mRNA following drug exposure. CONCLUSION: The present data suggested that heparin and staurosporine appears to interact in a synergistic manner, which may be regulated by the overexpression of c-myc, bax.更多还原