摘要
目的应用RNA干扰技术(RNA interference,RNAi),构建针对细胞周期素D1(Cyclin D1)的RNAi,并检测其对K562细胞增殖和对化疗药物敏感性的影响。方法采用针对Cyclin D1的RNAi质粒载体转染K562细胞,用反转录-聚合酶链反应和Western blot法检测Cyclin D1RNAi后mRNA和蛋白水平的表达;用MTT实验测定细胞的增殖;流式细胞术测定细胞周期和凋亡。结果针对Cyclin D1 RNAi转染白血病K562细胞后,可以明显降低白血病K562细胞中Cyclin D1的mRNA和蛋白的表达,明显抑制K562细胞的增殖,提高化疗药物Vp16诱导的细胞凋亡。结论利用RNAi抑制Cyclin D1可抑制白血病K562细胞增殖,提高对化疗药物Vp16的敏感性。
Objective To observe the gene silencing mediated by the specific RNAi targeted against Cyelin D1 and its effect on biological activity of human K562 leukemia cells. Methods RNAi plasmid vectors against Cyclin D1 were transfeeted into K562 leukemia cells. Cell activation was analysed by trypan blue reject test and cell proliferation inhibitions were determined by MTF assay. Cyclin D1 expressions were detected by reverse transcription - polymerase chain reaction （ RT - PCR） and western blot analysis. Cell cycle distribution and apoptosis was examined by flow cytometry. Results The RNAi vector targeted against Cyclin D1 efficiently suppressed Cyclin D1 expression at mRNA and protein levels. The RNAi - mediated gene silencing of Cyclin D1 resulted in significant inhibition of cell. Moreover, the cancer cells showed significant increased ratio of G1 phase, decreased ratio of S phase, and increased apoptosis induced by Vp16. Conclusion The specific RNAi targeted against Cyclin D1 could inhibit cell growth and increase sensitivity against Vp16 in K562 leukemia cells.
出处
《河北医科大学学报》
CAS
2012年第9期1008-1011,共4页
Journal of Hebei Medical University
