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目的 :利用bcl xS基因降低肿瘤细胞凋亡阈值 ,促进化疗药物顺铂诱导的鼻咽癌细胞凋亡 ,探索提高顺铂疗效的新途径。方法 :利用脂质体LipofectAmine将含有人bcl xS基因的重组真核表达质粒pcDNA3xS导入人鼻咽癌低分化上皮细胞株CNE 2Z ,G4 18筛选培养后 ,经Westernblot检测bcl xS的表达。以不含有bcl xS基因的pcDNA3质粒转染CNE 2Z作为对照细胞(CNE 2Zneo)。顺铂处理 2种细胞后 ,台盼蓝计数法求得各自的IC50 ,流式细胞仪及荧光染色检测凋亡细胞百分比。结果 :Westernblot检测证实获得稳定表达bcl xS的细胞 (CNE 2ZxS) ,与对照细胞相比 ,其对顺铂的IC50 值明显降低 ,经相同剂量顺铂处理后 ,流式细胞仪及荧光染色检测 ,结果表明 ,其凋亡细胞百分比明显高于对照细胞。结论 :bcl xS能显著增加鼻咽癌CNE 2Z细胞对顺铂诱导凋亡的敏感性
OBJECTIVE: To use bcl xS gene to reduce the apoptosis threshold of tumor cells and promote cisplatin-induced apoptosis of nasopharyngeal carcinoma cells and explore new ways to improve cisplatin efficacy. Methods: The recombinant eukaryotic expression plasmid pcDNA3xS containing human bcl xS gene was transfected into human nasopharyngeal carcinoma cell line NSE 2Z and G4 18 by liposome LipofectAmine. The expression of bcl xS was detected by Western blot. CNE 2Z was transfected as control cells (CNE 2Zneo) with pcDNA3 plasmid that does not contain the bcl xS gene. Cisplatin treatment of two kinds of cells, trypan blue counting their respective IC50, flow cytometry and fluorescent staining to detect the percentage of apoptotic cells. Results: Western blot analysis confirmed that cells with stable expression of bcl xS (CNE 2ZxS) had significantly lower IC50 values compared with control cells. After treatment with the same dose of cisplatin, the cells were detected by flow cytometry and fluorescent staining. The results showed that the percentage of apoptotic cells was significantly higher than the control cells. Conclusion: bcl xS can significantly increase the sensitivity of nasopharyngeal carcinoma CNE 2Z cells to cisplatin-induced apoptosis