摘要
背景与目的:实验证明bcl-xL在鼻咽癌CNE-2Z细胞中存在高表达,它可能在鼻咽癌的发生发展中发挥着重要的作用。本研究以脂质体(lipofectin)为载体,将人工合成的bcl-xL反义寡核苷酸(antisenseoligodeoxynucleotide,ASODN)片段导入CNE-2Z细胞,拟探讨ASODN对CNE-2Z细胞的影响。方法:以bcl-xL的编码区为靶点,人工合成20个碱基全硫代修饰的反义寡核苷酸,脂质体转染反义寡核苷酸进入CNE-2Z细胞后,用MTT法检测细胞成活率;用琼脂糖凝胶电泳、荧光染色、流式细胞术等方法检测细胞凋亡。结果:MTT结果发现,ASODN在Lip介导下即可显著抑制CNE-2Z细胞株细胞增殖(P<0.01),ASODN对细胞增殖的抑制作用随其浓度增加而增强;ASODN作用细胞36h后,经Hoechst33258/PI双染在荧光显微镜下可见细胞缩小、染色质固缩、核断裂;流式细胞仪分析发现在G1峰前出现一个亚二倍体峰即凋亡峰;琼脂糖凝胶电泳分析可见ASODN/Lip处理组出现明显的“梯状”外观等凋亡特征性改变。结论:ASODN脂质体转染CNE-2Z细胞后可促进CNE-2Z的细胞凋亡;在肿瘤研究中,bcl-xL可作为一个有用的靶基因,为开发鼻咽癌基因治疗药物提供实验依据。
BACKGROUND &OBJECTIVE:Recent studies have shown that overexpression of bcl-x L was detected in human nasopharyng eal carcinoma (NPC)cell strain CNE-2Z,sug g esting it may play a pivotal role in tumorig enesis of NPC.The current study was desig ne d to explore the effect of bcl-x L antisense olig odeoxynucleotide (ASODN)on CNE-2Z.METHODS :A 20-mer g apmer ASODN with a full phosphorothioate b ackbone targ eting a sequence unique of the bcl-x L coding reg ion was artificially synt hesized.Bcl-x L ASODN was transfected into CNE-2Z cells throu g h lipofectin.The survival rate was assessed by MTT assay and internucleosomal fr ag mentation of g enomic DNA was detec ted by ag arose g el electrophoresis.Apo ptotic chang es after treatment with ASODN were observed by fluorescence micro scopy and flow cytometry.RESULTS :MTT assay showed that the proliferation of CNE-2Z cells decreased sig nificantly after treatment with ASODN /Lip as compare d with control (P<0.01).ASODN /Lip reduced the proliferation of CNE-2Z in a dose-dependent manner.After tr eatment with ASODN /Lip for 36hours ,most cel ls stained with Hoechst 33258/PI exhibited apoptotic cell morpholog y such as cell shrinkag e,nuclear condensation,and nuclear frag ment ation under fluorescence microscop e;a apoptotic peak appeared on flow cyto metry;a ladder-like pattern of DNA frag mentation appeared on ag arose g el electrophoresis.CONCLUSION:ASODN can inhibit proliferation of CNE-2Z cells and induce apoptosis of CNE-2Z cells.The results sug g est that bcl-x_L is a promising targ et for g ene therap y of NPC.
出处
《癌症》
SCIE
CAS
CSCD
北大核心
2003年第1期11-15,共5页
Chinese Journal of Cancer
基金
广东省重点科技项目(2KM04205S)
