[Effect of CPEB4 on Proliferation and Apoptosis of Chronic Myeloid Leukemia Cells]

Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2019 Dec;27(6):1779-1785. doi: 10.19746/j.cnki.issn.1009-2137.2019.06.012.
[Article in Chinese]

Abstract

Objective: To explore the expression of CPEB4 in K562 cells, the biological activity and its possible molecular mechanisms.

Methods: Western blot was used to detect the expression of CPEB4 in normal leukocytes and K562 cells. The overexpression plasmid pcDNA3.1(+)-His-CPEB4 and the silencing plasmid pPLK+Puro-CPEB4 shRNA were transfected into K562 cells to change the expression of CPEB4 in K562 cells, and the transfection efficiency was detected by Western blot. Finally, CCK-8 and flow cytometry were used to detect the proliferation and apoptosis of differently treated cells, and the expression changes of proliferation and apoptosis marker proteins (AKT, p-AKT, caspase-3, BCL-2) were detected by Western blot.

Results: Compared with normal leukocytes, the expression of CPEB4 protein in K562 cells was higher (P<0.01). Compared with the control group, the proliferation of CPEB4-silenced K562 cells significantly increased (P<0.01), the number of apoptotic cell significantly decreased, and expression of AKT, p-AKT and BCL-2 was significantly increased, the protein expression of caspase-3 was significantly reduced. The proliferation of K562 cells after CPEB4 overexpression was slowed down (P<0.05), the number of apoptotic cells significantly increased,the expressions of AKT, p-AKT and BCL-2 were significantly down-regulated, and the expression of caspase-3 was up-regulated.

Conclusion: CPEB4 can inhibit the proliferation and promote the apoptosis of K562 cells, the AKT, p-AKT, BCL-2 and caspase-3 are involved in the regulation mechanism.

题目: CPEB4对慢性髓系白血病细胞增殖与凋亡的影响.

目的: 探索CPEB4在K562细胞中的表达、生物学活性及其可能的分子机制.

方法: 采用Western blot检测正常白细胞和K562细胞中CPEB4的表达情况;再将过表达质粒pcDNA3.1(+)-His-CPEB4、沉默质粒pPLK+Puro-CPEB4 shRNA电穿孔转染K562细胞,改变CPEB4在K562细胞中的表达量,应用Western blot检测转染效率;最后应用CCK-8和流式细胞术检测不同处理的细胞增殖和凋亡情况,并用Western blot法检测增殖与凋亡标志蛋白(AKT、p-AKT、caspase-3、BCL-2)的表达变化。.

结果: 与正常白细胞相比,K562细胞中CPEB4蛋白表达量较高(P<0.01);CPEB4沉默的K562细胞与对照组相比,细胞增殖水平显著增高(P<0.001),细胞凋亡数显著减少,AKT、p-AKT、BCL-2蛋白表达水平明显增高,而caspase-3蛋白表达显著降低;CPEB4过表达后的K562细胞增殖减慢(P<0.05),细胞凋亡数明显增加,AKT、p-AKT和BCL-2蛋白表达显著降低,caspase-3蛋白表达增高。.

结论: CPEB4可抑制K562细胞增殖,促进K562细胞凋亡,而AKT、p-AKT、BCL-2和Caspase-3等分子参与调控机制。.

MeSH terms

  • Apoptosis*
  • Cell Proliferation
  • Humans
  • K562 Cells
  • Leukemia, Myelogenous, Chronic, BCR-ABL Positive*
  • RNA-Binding Proteins / metabolism*
  • Transfection

Substances

  • CPEB4 protein, human
  • RNA-Binding Proteins