Effects of silencing of muscle blind-like protein 1 gene on cell proliferation, apoptosis and migration abilities of leukemia cell line K562
10.3760/cma.j.cn115356-20240320-00039
- VernacularTitle:沉默盲肌样蛋白1基因对白血病细胞株K562细胞增殖、凋亡及迁移能力的影响
- Author:
Jun WANG
1
;
Shenhua TANG
;
Cuicui WANG
;
Cong ZHU
;
Xiuhong JIA
Author Information
1. 滨州医学院附属医院儿科，滨州 256603
- Keywords:
Leukemia;
RNA interference;
Muscle blind-like protein 1;
Cell proliferation;
Apoptosis;
Cell migration assays
- From:
Journal of Leukemia & Lymphoma
2024;33(11):657-663
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the effects of targeted silencing of muscle blind-like protein 1 (MBNL1) gene on the proliferation, apoptosis and migration abilities of leukemia cell line K562 and their possible mechanisms.Methods:Single gene analysis was used to search for differences in MBNL1 gene expression between leukemia samples (173 cases) and healthy control samples (70 cases) in The Cancer Genome Atlas (TCGA) database. The data were updated in 2018. The logarithmic growth phase leukemia cell line K562 was taken and divided into sh-MBNL1 group (transfected with shRNA sequence with targeted silencing of MBNL1 gene), sh-NC group (transfected with corresponding negative control shRNA sequence) and blank control group (not transfected with shRNA sequence). Real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) was used to detect the relative expression levels of MBNL1, transforming growth factor β 1 (TGF-β 1) and Smad7 mRNA in each group of cells; Western blotting was used to detect the relative expression levels of cell migration-related proteins, apoptosis-related proteins, TGF-β 1, and Smad7 proteins; CCK-8 method was used to detect cell proliferation ability; Transwell method was used to detect cell migration ability. Results:In TCGA database, the relative expression level of MBNL1 gene in leukemia samples was higher than that in healthy control samples ( P < 0.05). The relative expression levels of MBNL1 protein in the sh-MBNL1 group, sh-NC group and blank control group were 0.71±0.11, 1.00±0.11 and 1.03±0.10, respectively, and the difference was statistically significant ( F = 7.78, P < 0.05); the relative expression level of MBNL1 protein in the sh-MBNL1 group was lower than that in the sh-NC group and blank control group (both P < 0.05). The results of CCK-8 assay showed that the cell proliferation ability of sh-MBNL1 group at 72 and 96 hours after transfection was higher than that of sh-NC group and blank control group (both P < 0.05). The Transwell method detection results showed that the number of cell membrane penetration in the sh-MBNL1 group, sh-NC group and blank control group were 666±135, 1 072±157 and 1 006±51, respectively, and the difference was statistically significant ( F = 9.40, P = 0.014); the number of cell membrane penetration in the sh-MBNL1 group was less than that in the sh-NC group and blank control group (both P < 0.05). The relative expression level of E-cadherin protein in the sh-MBNL1 group was higher than that in the sh-NC group and blank control group (both P < 0.01); the relative expression levels of Vimentin, Bax, caspase-3, TGF-β 1, and Smad7 proteins in the sh-MBNL1 group were lower than those in the sh-NC group and blank control group (all P < 0.01). The qRT-PCR detection results showed that the relative expression levels of TGF-β 1 mRNA and Smad7 mRNA in the sh-MBNL1 group were lower than those in the sh-NC group and blank control group (both P < 0.05). Conclusions:Silencing of MBNL1 gene can promote the proliferation of leukemia cell line K562, weaken its migration ability, and affect cell apoptosis. The mechanism may be related to the regulatory effect of TGF-β-Smad signaling pathway.
