Lipofectin介导bcl-xL反义寡核苷酸抑制鼻咽癌细胞增殖活性的研究

目的　观察并探讨阳离子脂质体 (L ipofectin,L ip)介导的 bcl- x L 反义寡核苷酸 (antisenseoligodeoxynucletide,ASODN)对鼻咽癌细胞株 CNE- 1、CNE- 2 Z细胞体外抑制增殖活性的影响。方法　人工合成硫代修饰型 bcl- x L 反义寡核苷酸 (ASODN) ,与 L ip混合制备成 L ip+oligodeoxynucletide(ODN)复合物 ,处理鼻咽癌细胞株 CNE- 1、CNE- 2 Z细胞 ,用 MTT比色法检测细胞存活状态 ;流式细胞术检测细胞凋亡。结果　 ASODN处理细胞 3 6小时 ,于 40 0 μm ol/ L 浓度条件下鼻咽癌细胞的增殖受到明显抑制 ,且这一抑制作用随着浓度的增加与时间的延长而加强 ,与硫代修饰型随机序列对照寡核苷酸 (sequence control ODN,SCODN)组及单独脂质体组比较 ,差异有显著性 (P<0 .0 5 )。结论　 bcl- x L 反义寡核苷酸可显著抑制鼻咽癌细胞株 CNE- 1、CNE- 2 Z细胞体外增殖活性 ,其抑制作用可能通过封闭 bcl- x L 基因表达实现 ,bcl- x L 反义寡核苷酸有可能成为一种潜在性的基因治疗药物用于鼻咽癌的治疗

Study on proliferation inhibition of nasopharyngeal carcinoma cells by Lipofecton-mediated bcl-xL antisense oligodeoxynucleotides

Objective To investigate the effect on proliferation inhibition of human nasopharyngeal carcinoma (NPC) cells by Lipofectin mediated bcl xL antisense oligodeoxynucleotide (ASODN).Methods ASODN and sequence control ODN (SCODN) were synthesized and combined to Lipofectin (Lip) to prepare Lip+oligonucleotides(ODN) complexes. After exposure to Lip+ODN complexes for 36 hours and continuous incubation in complete medium for another 24 hours, the growth inhibition of human NPC cell lines CNE 1 and CNE 2Z was measured by MTT assay and flow cytometry(FCM) was used to observe apoptotic changes in ASODN treated cells.Results Treatment of CNE 1, CNE 2Z cells with ASODN at a concentration of 400 nmol/L for 36 hours significantly decreased the viability. The effect of inhibition of cells was enhanced with increasing concentration and exposure time. This resulted in the induction of apoptosis characterized by apoptotic peak. Cells incubated with the sequence control ODN(SCODN) or Lipofectin alone had no similar effect. Conclusions Lipofectin mediated bcl xL ASODN could efficiently inhibit cell growth of NPC cell lines CNE 1 and CNE 2Z.ASODN may be used in the gene therapy for human nasopharyngeal carcinoma.