Department of Hematology, Affiliated Hospital of Nantong University, Nantong, Jiangsu, China.
Department of Oncology, Nantong Oncology Hospital, Nantong, Jiangsu, China.
Cell Transplant. 2021 Jan-Dec;30:963689720980367. doi: 10.1177/0963689720980367.
The present study aimed to investigate the effect and possible mechanism of recombinant human thrombopoietin (rhTPO) on mouse 32D cells (a mouse myeloid progenitor cell line) treated with serum from patients with aplastic anemia and to elucidate the potential mechanism of rhTPO in the treatment of aplastic anemia. After treatment with aplastic anemia serum, the apoptotic rate of 32D cells was increased and the proliferation of 32D cells was significantly inhibited. rhTPO reduced the apoptotic rate and promoted the proliferation of 32D cells, while rhTPO failed to restore the cell proliferation of 32D cells from aplastic anemia serum group to the normal level as compared to that from the normal serum group. The phosphorylation level of STAT3 protein was higher, and the phosphorylation level of STAT5 protein was lower in 32D cells from aplastic anemia serum group than that in normal serum group. After rhTPO treatment, the phosphorylation level of STAT3 protein in aplastic anemia serum group was decreased and the phosphorylation level of STAT5 protein was increased. The expression levels of Survivin and Bcl-2 were significantly decreased in 32D cells from aplastic anemia serum group, which were significantly increased after rhTPO treatment. The expression level of Bax protein in 32D cells from the normal serum group after rhTPO treatment was significantly decreased; while the mRNA expression level of Bax was not affected by rhTPO. The expression levels of Bax mRNA and protein were significantly up-regulated in 32D cells from aplastic anemia serum group, which was significantly decreased by rhTPO treatment. In conclusion, our results indicated that aplastic anemia serum impaired proliferative potential and enhanced apoptosis of 32D cells. Further mechanistic studies revealed that rhTPO promoted cell proliferation and attenuated apoptosis of aplastic anemia serum-treated 32D cells via activating STAT3/STAT5 signaling pathway and modulating apoptosis-related mediators.
本研究旨在探讨重组人血小板生成素（rhTPO）对贫血患者血清处理的小鼠 32D 细胞（一种鼠髓系祖细胞系）的作用及其可能机制，并阐明 rhTPO 治疗再生障碍性贫血的潜在机制。贫血患者血清处理后，32D 细胞的凋亡率增加，细胞增殖明显受到抑制。rhTPO 降低 32D 细胞的凋亡率，促进细胞增殖，但与正常血清组相比，rhTPO 未能将贫血血清组 32D 细胞的细胞增殖恢复到正常水平。贫血血清组 32D 细胞中 STAT3 蛋白磷酸化水平升高，STAT5 蛋白磷酸化水平降低。rhTPO 处理后，贫血血清组 32D 细胞中 STAT3 蛋白磷酸化水平降低，STAT5 蛋白磷酸化水平升高。贫血血清组 32D 细胞中 Survivin 和 Bcl-2 的表达水平明显降低，rhTPO 处理后明显增加。rhTPO 处理后，正常血清组 32D 细胞 Bax 蛋白表达水平明显降低；而 rhTPO 对 Bax 的 mRNA 表达水平无影响。rhTPO 处理后，贫血血清组 32D 细胞 Bax mRNA 和蛋白表达水平明显上调。综上所述，本研究结果表明贫血患者血清可损害 32D 细胞的增殖潜能，增强其凋亡。进一步的机制研究表明，rhTPO 通过激活 STAT3/STAT5 信号通路和调节凋亡相关介质，促进细胞增殖，减轻贫血患者血清处理的 32D 细胞凋亡。
