RNAi干扰对K562细胞表达谱的影响

Study on variety of gene expression profiling in K562 cells by RNAi

目的联合运用RNAi技术和cDNA表达谱芯片研究干扰c-myc对K562细胞表达谱的影响。方法经过RT-PCR和流式细胞仪检测,筛选出干扰效果最好的siRNAs,经LipofectamineTM2000脂质体法转染K562细胞,提取总RNA,逆转录为cDNA,荧光标记后与K562表达谱芯片杂交。结果经扫描、分析杂交结果,有455个基因表达下调,有12个基因表达上调。结论基因芯片和RNAi是分析基因功能的有力工具,也是发现新的肿瘤治疗靶标的有效方法。

Objective To study the changes of gene expression profile in K562 cells which were treated by RNA interference method targeting c-myc. Methods siRNAs were designed and functionally selected with RT-PCR and FACS. The siRNA targeting e-myc was indentificd and transfected the cultured K562 cells in vitro with lipofectamineTM 2000. The total RNA were extracted and reverse transcribed into cDNA,and hybridized to the K562 cDNA microarray after cy5- and cy3-1abeling. Results The hybridized microarray was scanned. The analysis for the data revealed that there were 455 down-regulated genes and 12 up-regulated genes. Conclusion RNAi coupled with microarray analysis may provide an efficient system to define the role of specific genes and a novel therapeutic tool for cancer.