Wang Nan, Lyu Na, Min Xin, Wang Li-Li, Zhu Hai-Yan
Department of Hematology, The First Medical Center, Chinese PLA General Hospital, Beijing 100853, China.
Department of Hematology, The First Medical Center, Chinese PLA General Hospital, Beijing 100853, China,E-mail:
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2021 Apr;29(2):389-394. doi: 10.19746/j.cnki.issn.1009-2137.2021.02.013.
To investigate the effects of recombinant human thrombopoietin (rhTPO) to proliferation and apoptosis of acute myeloid leukemia (AML) cell lines.
After the treatment of different concentrations of rhTPO (0, 50, 100 ng/ml) for different time (24,48,72 h)，the cell proliferation rates of the AML cell lines (Kasumi-1, Skno-1, HEL, HL-60, THP-1) were determined by CCK-8 method. Apoptosis rate of each cell line cocultured with rhTPO was detected by Annexin V/PI method. The relative expression of TPO receptor c-MPL (myeloproliferative clonal antibody) mRNA in AML cell lines was detected by Q-PCR. The expression of c-MPL protein in each cell line was detected by Western blot. The expression of c-MPL antigen in HL-60 cells treated by different concentrations of rhTPO was detected by Flow cytometry.
RhTPO showed no promotion to the proliferation of Kasumi-1, Skno-1, HEL, HL-60, THP-1 cell lines，however，it showed inhibitory effect to cell proliferation (72 h 0 ng/ml vs 100 ng/ml, P= 0.029) and pro-apoptotic (48 h 0 ng/ml vs 50 ng/ml, P=0.0143) in HL-60 cells. In Kasumi-1, Skno-1, HEL and THP-1 cells, there showed no statistically significant differences in apoptosis rate among each groups treated by different concentrations of rhTPO. Each AML cell line showed different levels of c-MPL gene and c-MPL protein expression, but HEL cells showed the highest expression in both of them. After HL-60 cells were treated by different concentrations of rhTPO for 48 hours, there showed no statistical difference in c-MPL antigen expression among each groups.
RhTPO can not promote the proliferation of Kasumi-1, Skno-1, HEL, HL-60 and THP-1 leukemia cell lines. On the contrary, rhTPO can inhibit HL-60 cell proliferation and promote its apoptosis, and this effect is not related to c-MPL gene expression or protein expression.
探讨重组人血小板生成素（rhTPO）对急性髓系白血病（AML）细胞系增殖和凋亡的影响。
用不同浓度的rhTPO（0、50、100 ng/ml）处理不同时间（24、48、72小时）后，采用CCK-8法检测AML细胞系（Kasumi-1、Skno-1、HEL、HL-60、THP-1）的细胞增殖率。采用Annexin V/PI法检测与rhTPO共培养的各细胞系的凋亡率。采用Q-PCR检测AML细胞系中TPO受体c-MPL（骨髓增殖性克隆抗体）mRNA的相对表达。采用Western blot检测各细胞系中c-MPL蛋白的表达。采用流式细胞术检测不同浓度rhTPO处理的HL-60细胞中c-MPL抗原的表达。
rhTPO对Kasumi-1、Skno-1、HEL、HL-60、THP-1细胞系的增殖无促进作用，然而，其对HL-60细胞的增殖有抑制作用（72小时，0 ng/ml与100 ng/ml相比，P = 0.029），并具有促凋亡作用（48小时，0 ng/ml与50 ng/ml相比，P = 0.0143）。在Kasumi-1、Skno-1、HEL和THP-1细胞中，不同浓度rhTPO处理的各组之间凋亡率无统计学差异。各AML细胞系的c-MPL基因和c-MPL蛋白表达水平不同，但HEL细胞在两者中表达最高。HL-60细胞经不同浓度rhTPO处理48小时后，各组之间c-MPL抗原表达无统计学差异。
rhTPO不能促进Kasumi-1、Skno-1、HEL、HL-60和THP-1白血病细胞系的增殖。相反，rhTPO可抑制HL-60细胞增殖并促进其凋亡，且这种作用与c-MPL基因表达或蛋白表达无关。
