小RNA干扰蛋白酶体亚基α7抑制K562细胞增殖  

Knockdown of Proteasome Subunit α7 with Small Interfering RNA Inhibits Cell Proliferation of K562 Cell Line

作　　者：秦涛[1,2] 范翠青[1,2] 朱宁[1,2] 沈岩[1,2] 陈梅红[1,2] 

机构地区：[1]中国医学科学院基础医学研究所 [2]北京协和医学院基础学院生物化学与分子生物学系医学分子生物学国家重点实验室,北京100005

出　　处：《中国医学科学院学报》2013年第6期601-606,共6页Acta Academiae Medicinae Sinicae

基　　金：国家自然科学基金(31050008)~~

摘　　要：目的运用小干扰RNA(siRNA)技术降低人髓性白血病细胞株K562细胞20S蛋白酶体亚基α7(PSMA7)基因的表达水平,探讨PSMA7表达下调对其细胞功能的影响。方法采用HiPerFect将化学合成的针对人PSMA7基因的特异性siRNA转染入K562细胞后,用Western blot法检测siRNA对PSMA7蛋白表达的抑制效果,采用MTS法及细胞计数法检测细胞增殖速率,流式细胞术检测细胞周期分布,Western blot法检测细胞周期蛋白Cyclin A、D、E的表达水平,Annexin V/FITC染色后流式细胞术检测细胞凋亡率。结果 PSMA7 siRNA转染K562细胞后使PSMA7蛋白表达水平明显下调;细胞增殖速率明显降低但无明显细胞凋亡,细胞周期中S期细胞比例减少;细胞周期蛋白Cyclin A、E表达下调。结论 PSMA7表达下调可使人髓性白血病细胞株K562细胞Cyclin A、E表达下调,使S期细胞比例降低从而抑制K562细胞增殖。Objective To study the effect of human proteasome subunit α7 （PSMA7） gene silencing by small interfering RNA （siRNA） on human myeloid leukemia cell line K562. Methods PSMA7 gene-specific siRNA was chemically synthesized and transfected into K562 cell line by HiPerFect. The expression level of PSMA7 protein was detected by Western blot analysis. Cell proliferation was determined by MTS and cell count- ing. Cell cycle distribution was measured by flow cytometry. The expressions of Cyclin A, D, and E were detec- ted by Western blot analysis. The apoptotic ratio was determined by flow cytometry. Results PSMA7 protein was evidently silenced 48 hours after transfection of the PSMA7-specific siRNA into K562 cell line. The proliferation of the cells was markedly inhibited, and the percentage of S phase cells decreased. However, no apoptosis was observed. The expressions of Cyclin A and E were down-regulated. Conclusion Knockdown of PSMA7 down-reg- ulates the expression of Cyclin A and E and thus decreases the proportion of cells in S phase; as a result, the proliferation of K562 cell line is inhibited.

关 键 词：RNA干扰 蛋白酶体亚基α7 细胞周期蛋白 细胞增殖 

分 类 号：Q751[生物学—分子生物学]