Objective: To construct a K562 and adriamycin-resistant K562 (KAR) cell line with stably down-regulation of NCL (nucleolin) expression, and to investigate the effect of NCL down-regulation on the drug resistance in K562 and KAR cells.
Methods: K562 and KAR cells were infected with lentivirus, and stably transfected cell clones were obtained by puromycin screening. The cell proliferation was detected by MTS assay, the cell apoptosis was detected by flow cytometry, and the expression level of drug resistance related genes was detected by real-time PCR.
Results: The K562 and KAR cells with stable down-regulation of NCL were successfully constructed. Compared with the control group, the proliferation of K562 and KAR cells with down-regulating NCL expression decreased significantly (P ＜0.05), the apoptosis of cells increased significantly (P ＜0.05), and cell resistance to adriamycin was down-regulated.
Conclusion: Inhibition of NCL expression may increase the sensitivity of cells to adriamycin, which may be related with the promotion of apoptosis of K562 and KAR cells.
题目: 抑制NCL表达对慢性髓系白血病细胞K562及其耐药细胞的影响.
目的: 建立稳定下调核仁素（NCL）表达的K562细胞及耐阿霉素的K562细胞（KAR），研究下调NCL对K562及KAR细胞耐药作用的影响.
方法: 用慢病毒感染K562及KAR细胞，经嘌呤霉素筛选获得稳定转染的细胞克隆。采用MTS法检测各组细胞的增殖能力，流式细胞术检测细胞凋亡情况，Real-time PCR检测耐药相关基因的表达水平.
结果: 成功的构建了NCL表达下调的K562及KAR稳定转染细胞株。与对照组相比较，下调NCL的表达可以使细胞的增殖明显受抑（P＜0.05），细胞的凋亡显著增加（P＜0.05），细胞对阿霉素的耐药性降低.
结论: 抑制NCL的表达可以提高细胞对阿霉素的敏感性，这一作用可能与其促进细胞凋亡有关.