[1]岳光星,张明智,张旭东,等.shRNA干扰沉默Mcl-1基因对淋巴瘤细胞系增殖的影响[J].第三军医大学学报,2012,34(12):1210-1213.
 Yue Guangxing,Zhang Mingzhi,Zhang Xudong,et al.Effect of shRNA interference targeting myeloid leukemia-1 gene on proliferation in lymphoma cell line[J].J Third Mil Med Univ,2012,34(12):1210-1213.
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
34卷
期数:
2012年第12期
页码:
1210-1213
栏目:
论著
出版日期:
2012-06-30

文章信息/Info

Title:
Effect of shRNA interference targeting myeloid leukemia-1 gene on proliferation in lymphoma cell line
作者:
岳光星张明智张旭东许伟朋陈新峰陈昱丞曹玲杨黎盛誉乔
郑州大学第一附属医院:肿瘤,骨科;河南省高等学校临床医学重点学科开放实验室
Author(s):
Yue Guangxing Zhang Mingzhi Zhang Xudong Xu Weipeng Chen Xinfeng Chen Yucheng Cao Ling Yang Li Sheng Yuqiao
Department of Ooncology, Department of Orthopedics, Open Laboratory of Key Disciplines in Clinical Medicine,  First Affiliated Hospital,  Zhengzhou University, Zhengzhou, Henan Province, 450052, China
关键词:
淋巴瘤髓细胞白血病基因-1短发夹RNA凋亡慢病毒载体
Keywords:
lymphoma Mcl-1 shRNA apoptosis lentiviral vector
分类号:
R394-33;R730-362;R733
文献标志码:
A
摘要:
目的      研究慢病毒介导的髓细胞白血病基因(Mcl-1)短发夹状RNA(shRNA)干扰对淋巴瘤细胞系SNK-6增殖和凋亡的影响。      方法      设计以Mcl-1为靶点的shRNA并构建携带此shRNA的慢病毒载体,转染淋巴瘤细胞系SNK-6,荧光定量PCR(qPCR)及Western blot方法检测病毒感染前后Mcl-1 mRNA及蛋白表达变化,四甲基偶氮唑盐法(MTT法)和流式细胞仪分析细胞增殖和凋亡变化的情况。      结果      成功构建Mcl-1-shRNA慢病毒表达载体,并可有效感染淋巴瘤细胞株SNK-6,感染后SNK-6细胞 Mcl-1基因的mRNA水平及蛋白水平表达明显下调,MTT法检测显示慢病毒介导的Mcl-1基因shRNA干扰与对照病毒组相比可明显抑制SNK-6细胞增殖[(31.6±3.3)%  vs (5.8±2.7)%, P<0.01], 流式细胞仪测定感染后细胞凋亡明显高于对照病毒组[(28.9±2.1)% vs (5.3±1.5)%, P<0.01]。       结论      慢病毒介导的Mcl-1基因shRNA干扰技术可特异性阻断SNK-6细胞Mcl-1基因的表达,抑制细胞的增殖并促进细胞的凋亡。
Abstract:
Objective      To explore the effect of lentivirus-mediated short hairpin RNA (shRNA) interference  targeting myeloid leukemia-1 (Mcl-1) gene on the proliferation and apoptosis of lymphoma cell line SNK-6.       Methods      The shRNA targeting Mcl-1 gene was designed and the lentiviral vector carrying the shRNA was constructed. Lymphoma SNK-6 cells were transfected with the recombinant lentiviral vectors (shRNA group) and the blank lentiviral vectors (control group), respectively. Real-time PCR and Western blotting were used to analyze the mRNA and protein expression of Mcl-1 gene in SNK-6 cells before and after transfection. MTT assay and flow cytometry were applied to detect the proliferation and apoptosis of the transfected SNK-6 cells, respectively.        Results      The shRNA targeting Mcl-1 gene was successfully inserted into the lentiviral vectors, which could effectively infect lymphoma SNK-6 cells. The mRNA and protein expression of Mcl-1 gene in the shRNA group significantly decreased as compared with those in the control group. The MTT assay results suggested that the proliferation of SNK-6 cells was significantly inhibited in the shRNA group as compared with that in the control group [(31.6±3.3)% vs (5.8±2.7)%, P<0.01]. The flow cytometry showed that the apoptosis rate of the shRNA group was significantly higher than that of the control group [(28.9±2.1)% vs (5.3±1.5)%, P<0.01].       Conclusion      Lentivirus-mediated shRNA interference targeting Mcl-1 gene can specifically block Mcl-1 gene expression, and inhibit the proliferation and promote the apoptosis of SNK-6 cells.

参考文献/References:

岳光星, 张明智, 张旭东, 等. shRNA干扰沉默Mcl-1基因对淋巴瘤细胞系增殖的影响[J].第三军医大学学报,2012,34(12):1210-1213.

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更新日期/Last Update: 2012-06-15