论文部分内容阅读
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 & AIM: Bcl-xL is highly expressed in nasopharyngeal carcinoma CNE-2Z cells. It may play an important role in the development of nasopharyngeal carcinoma. In this study, a synthetic bcl-xL antisense oligodeoxynucleotide (ASODN) fragment was introduced into CNE-2Z cells using lipofectin as a carrier to investigate the effect of ASODN on CNE-2Z cells. Methods: The antisense oligonucleotide modified by 20 bases per-thio was synthetized by targeting the coding region of bcl-xL. The antisense oligonucleotide was transfected into CNE-2Z cells with Cell viability was detected by MTT assay. Apoptosis was detected by agarose gel electrophoresis, fluorescence staining and flow cytometry. Results: MTT results showed that ASODN could significantly inhibit the proliferation of CNE-2Z cells (P <0.01) under the mediation of Lip, the inhibitory effect of ASODN on cell proliferation increased with the concentration of ASODN; Hoechst33258 / PI double staining under the fluorescence microscope shows cell shrinkage, chromatin condensation, nuclear rupture; flow cytometry analysis showed a sub-diploid peak in the G1 peak before apoptosis peak; agarose gel electrophoresis analysis can be seen ASODN / Lip treatment group showed obvious “ladder” appearance and other apoptosis characteristic changes. CONCLUSION: ASODN liposome transfected CNE-2Z cells can promote CNE-2Z cell apoptosis. In tumor research, bcl-xL can be used as a useful target gene to provide experimental evidence for the development of gene therapy for nasopharyngeal carcinoma .