Department of Hematology, The First Affiliated Hospital of Wannan Medical College (Yijishan Hospital of Wannan Medical College), Wuhu City, 241001, Anhui Province, China.
Wannan Medical College, Wuhu City, 241001, Anhui Province, China.
Biotechnol Lett. 2020 Jul;42(7):1275-1286. doi: 10.1007/s10529-020-02878-1. Epub 2020 Mar 31.
To investigate the functions of eIF3b in chronic myelogenous leukemia (CML).
The expression of eIF3b was inhibited by transfecting aspecifically designed shRNA into the CML cell lines of TK-6 and K562. The CCK8 assay was conducted to determine cell viability, and flow cytometry was used to examine the change in the cell cycle and cell apoptosis. RNAsequencing was applied to screen the candidate targets of eIF3b to identify the underlying mechanisms of eIF3b.An in vivo tumour xenograft mouse model was established by injecting shRNA transfected cells into the NCG mice. The tumour size and body weight of mice were monitored every other day. The mice were sacrificed 2 weeks after the tumour cell injection. The expression of eIF3b and target genes in the tumour tissues were determined by immunohistochemical staining and Western blotting.
The group with inhibited expression of eIF3b led to about 50% lower cell viability compared with that of the control group (P < 0.05). Flow cytometry suggested that the percentage of increase in apoptotic cells was eight times higher than those in control group for TK-6 and K562 cells (P < 0.05). However, the difference between the cell amounts in the S phase for the experiment and control groups was not significant. After RNAsequencing and further validation via qPCR, C3G was screened as the potential target of eIF3b involved in the cell proliferation and apoptosis of CML cell lines. Subsequent in vivo analysis proved that the inhibition of eIF3b suppressed tumour formation and decreased C3G expression, thereby indicating that C3G was the potential target of eIF3b.
eIF3b is correlated with the cell proliferation and cell apoptosis of CML. Moreover, eIF3b regulation most probably occurs via regulating the expression of C3G.
研究 eIF3b 在慢性髓系白血病（CML）中的作用。
通过转染特异性设计的 shRNA 抑制 CML 细胞系 TK-6 和 K562 中的 eIF3b 表达。通过 CCK8 测定法测定细胞活力，通过流式细胞术检测细胞周期和细胞凋亡的变化。应用 RNA 测序筛选 eIF3b 的候选靶标，以确定 eIF3b 的潜在作用机制。通过将转染 shRNA 的细胞注射到 NCG 小鼠中建立体内肿瘤异种移植小鼠模型。每隔一天监测小鼠的肿瘤大小和体重。在肿瘤细胞注射后 2 周处死小鼠。通过免疫组织化学染色和 Western blot 测定肿瘤组织中 eIF3b 和靶基因的表达。
与对照组相比，eIF3b 表达受抑制的组细胞活力降低约 50%（P < 0.05）。流式细胞术表明，与对照组相比，TK-6 和 K562 细胞中凋亡细胞的百分比增加了 8 倍（P < 0.05）。然而，实验组和对照组之间 S 期细胞数量的差异并不显著。经过 RNA 测序和进一步通过 qPCR 验证，筛选出 C3G 是 eIF3b 参与 CML 细胞系细胞增殖和凋亡的潜在靶标。随后的体内分析证明，抑制 eIF3b 抑制肿瘤形成并降低 C3G 表达，表明 C3G 是 eIF3b 的潜在靶标。
eIF3b 与 CML 的细胞增殖和细胞凋亡有关。此外，eIF3b 的调节可能是通过调节 C3G 的表达来实现的。
