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目的:探讨双氢青蒿素(DHA)对下咽癌Fadu细胞凋亡的影响。方法:用不同浓度DHA干预下咽癌Fadu细胞,观察DHA对细胞形态的影响;分别于24h或48h后用MTT法检测细胞增殖的变化并计算IC50值。用AnnexinⅤ-FITC凋亡检测试剂盒分析DHA对Fadu细胞凋亡的影响。用Western Blot法测定DHA对Bcl-2、Bcl-xl、Mcl-1、Bax和C-PARP凋亡相关基因的影响。结果:一定浓度的DHA可显著改变细胞的形态学变化,抑制下咽癌Fadu细胞增殖且明显增加凋亡细胞的数量,该变化与干预浓度和时间呈依赖关系。进一步研究结果提示,DHA可显著抑制Bcl-2、Bcl-xl和Mcl-1抗凋亡蛋白的表达,同时增加促凋亡蛋白Bax和C-PARP的表达。结论:DHA可显著抑制下咽鳞状细胞癌Fadu细胞的增殖,并明显增加细胞的凋亡,其机制与DHA调控Bcl-2家族密切相关。提示,Bcl-2家族成员在DHA抑制下咽鳞状细胞癌Fadu细胞增殖的过程中发挥了重要作用。
Objective: To investigate the effect of dihydroartemisinin (DHA) on the apoptosis of hypopharyngeal carcinoma Fadu cells. Methods: The effect of DHA on the cell morphology was observed with the intervention of DHA at different concentrations. The cell proliferation was measured by MTT assay at 24 or 48 hours, and the IC50 values were calculated. Annexin V-FITC apoptosis detection kit was used to analyze the effect of DHA on Fadu cell apoptosis. The effects of DHA on the apoptosis-related genes of Bcl-2, Bcl-xl, Mcl-1, Bax and C-PARP were determined by Western Blot. Results: A certain concentration of DHA could significantly change the morphological changes of cells, inhibit the proliferation of hypopharyngeal carcinoma Fadu cells and significantly increase the number of apoptotic cells, the changes with the intervention concentration and time-dependent. Further studies suggest that DHA can significantly inhibit the expression of Bcl-2, Bcl-xl and Mcl-1 anti-apoptotic protein, while increasing the expression of pro-apoptotic protein Bax and C-PARP. Conclusion: DHA can significantly inhibit the proliferation of Fadu cells and significantly increase the apoptosis of hypopharyngeal squamous cell carcinoma. The mechanism is closely related to DHA-regulated Bcl-2 family. These results suggest that Bcl-2 family members play an important role in DHA-induced proliferation of Fadu cells.