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目的检测胃癌细胞系中端粒酶RNA干涉后是否改变其他基因的表达。方法根据RNA干涉原理,针对端粒酶基因序列设计了抑制端粒酶表达的核苷酸序列,克隆入真核表达载体中;并以脂质体转入到胃癌细胞系的MGC80-3和BGC-823细胞中,以空质粒为对照,48 h后收集细胞的总RNA,逆转录成cDNA后,通过荧光差异显示法检测基因表达的变化。结果胃癌细胞BGC-823细胞和MGC80-3细胞中一些基因的表达量明显下降。结论荧光差异显示法能比较系统地检测与端粒酶活性相关的基因,为进一步研究这些基因与端粒酶之间的表达调控方式及其对细胞生长的影响打下了基础。
Objective To determine whether the expression of other genes is altered after telomerase RNA interference in gastric cancer cell lines. Methods Based on the principle of RNA interference, the nucleotide sequence of telomerase inhibition was designed according to the telomerase gene sequence and cloned into a eukaryotic expression vector. The liposome was transfected into MGC80-3 and BGC cells. In the -823 cells, empty plasmids were used as controls. After 48 hours, the total RNA of the cells was collected and reverse transcribed into cDNA. The changes in gene expression were detected by fluorescence differential display. Results The expression levels of some genes in gastric cancer cells BGC-823 cells and MGC80-3 cells were significantly decreased. Conclusion The fluorescence difference display method can systematically detect the genes related to telomerase activity, which lays the foundation for further study on the expression regulation mode of these genes and telomerase and its effect on cell growth.