天津医药

天津医药 ›› 2016, Vol. 44 ›› Issue (6): 679-682.doi: 10.11958/20150299

• 细胞与分子生物学 • 上一篇    下一篇

RNAi 靶向沉默 CDX2 基因对白血病细胞生物学行为的影响及其作用机制

孙维梅 1， 李建厂 1， 贾秀红 1， 李有杰 2， 唐慎华 1   

  1. 1山东滨州，滨州医学院附属医院儿科 （邮编 256603）； 2滨州医学院肿瘤分子生物学重点实验室
  • 收稿日期:2015-11-09 修回日期:2016-01-25 出版日期:2016-06-15 发布日期:2016-07-04
  • 基金资助:
    ： 山东省医药卫生科技发展计划 （2014WS0183）； 山东省自然科学基金 （ZR2014HL032）

Effects of RNAi targeting CDX2 gene expression on biological characters of human leukemia cells and its relevant mechanisms

SUN Weimei1, LI Jianchang1, JIA Xiuhong1, LI Youjie2, TANG Shenhua1   

  1. 1 Department of Pediatrics, Binzhou Medical University Hospital, Binzhou 256603, China;2 Department of Biochemistry and Molecular Biology, Binzhou Medical University
  • Received:2015-11-09 Revised:2016-01-25 Published:2016-06-15 Online:2016-07-04

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摘要： Abstract: Objective To detect the effects of siRNA targeting CDX2 gene expression on of BCR-ABL, caspase and Bax expressions, and the mechanisms thereof. Methods According to the earlier experiments, siRNA specifically targeting CDX2 gene (CDX2-siRNA) and the negative control sequence (CDX2-siRNA-NC) were selected, and then were transfected into K562 cells by Roche X-tremeGENE HP DNA Transfection Reagent. The flow cytometry analysis was used to detect the effects of siRNA on cell apoptosis. The expressions of BCR-ABL, caspase- 9, Bax mRNA and protein were tested by RTPCR and Western blot assay. Results MTT and flow cytometry analysis showed that after the silence of CDX2 gene expression, the proliferation of K562 cells was prohibited and the apoptotic rate of K562 cells was distinctly increased compared with that of normal cell group, but the negative control group had no significant change. According to the RT-PCR and Western blot assay, in comparison with the normal cell group and the negative control group, the expression levels of BCR-ABL mRNA and protein were obviously decreased, and the difference was statistic significance. On the other hand, the expressions of caspase- 9 and Bax mRNA and protein were significantly higher than those of other two groups (P＜0.05).Conclusion CDX2-siRNA can promote apoptosis of K562 cells obviously, and the mechanism is related with the downregulation of BCR-ABL and the up-regulation of caspase-9 and Bax.

关键词: 白血病, RNA 干扰, 基因沉默, 细胞凋亡, 融合蛋白质类, BCR-ABL, CDX2, RNA 干扰, 半胱氨酸天冬氨酸蛋白酶-9； Bax

Abstract: Abstract: Objective To detect the effects of siRNA targeting CDX2 gene expression on of BCR-ABL, caspase and Bax expressions, and the mechanisms thereof. Methods According to the earlier experiments, siRNA specifically targeting CDX2 gene (CDX2-siRNA) and the negative control sequence (CDX2-siRNA-NC) were selected, and then were transfected into K562 cells by Roche X-tremeGENE HP DNA Transfection Reagent. The flow cytometry analysis was used to detect the effects of siRNA on cell apoptosis. The expressions of BCR-ABL, caspase- 9, Bax mRNA and protein were tested by RTPCR and Western blot assay. Results MTT and flow cytometry analysis showed that after the silence of CDX2 gene expression, the proliferation of K562 cells was prohibited and the apoptotic rate of K562 cells was distinctly increased compared with that of normal cell group, but the negative control group had no significant change. According to the RT-PCR and Western blot assay, in comparison with the normal cell group and the negative control group, the expression levels of BCR-ABL mRNA and protein were obviously decreased, and the difference was statistic significance. On the other hand, the expressions of caspase- 9 and Bax mRNA and protein were significantly higher than those of other two groups (P＜0.05). Conclusion CDX2-siRNA can promote apoptosis of K562 cells obviously, and the mechanism is related with the downregulation of BCR-ABL and the up-regulation of caspase-9 and Bax.

Key words: leukemia, RNA interference, gene silencing, apoptosis, fusion proteins, BCR-ABL, CDX2, RNA interference, Caspase-9, Bax