中文摘要：

目的探讨膜结合型巨噬细胞集落刺激因子（mM-CSF）及其剪切体（mM-CSF-Δ）对淋巴细胞白血病Ramos细胞增殖的抑制作用。方法采用Overlap PCR法构建带有m M-CSF的真核表达质粒pTARGET-mM-CSF,再进一步构建胞内区截短30个氨基酸的表达质粒p TARGET-m M-CSF-Δ,并进行PCR及DNA双向测序鉴定。将空载体p TARGET、p TARGET-mM-CSF、pTARGET-mM-CSF-Δ质粒分别转染Ramos细胞,经G418筛选稳定表达细胞株,并用RT-PCR、Western blotting进行鉴定;MTT法检测细胞增殖,流式细胞仪检测细胞周期。结果成功构建了m M-CSF和m M-CSF-Δ的真核表达载体,获得了稳定转染细胞株Ramos-V、Ramos-M和Ramos-Δ。Ramos-M、Ramos-Δ、Ramos-V细胞增殖能力的OD值分别为0.413±0.014、0.384±0.019、0.463±0.037,Ramos-M细胞与Ramos-Δ细胞比较,Ramos-M、Ramos-Δ细胞分别与Ramos-V细胞比较,P〈0.05或〈0.01。Ramos-M、Ramos-Δ、Ramos-V细胞处于G0/G1期的比例分别为41.54%±1.22%、45.60%±1.09%、39.20%±1.53%,Ramos-M细胞与Ramos-Δ细胞比较,Ramos-M、Ramos-Δ细胞分别与Ramos-V细胞比较,P〈0.05或〈0.01。结论 mM-CSF、mM-CSF-Δ均能抑制淋巴细胞白血病Ramos细胞增殖,且后者抑制作用更强。

英文摘要：

Objective To investigate the inhibitory effect of membrane-bound macrophage colony-stimulating factor （ mM-CSF） and its spliceosome （ mM-CSF-Δ） on proliferation of lymphocytic leukemia cell line Ramos.Methods The eukaryotic expression plasmid of pTARGET-mM-CSF with mM-CSF was constructed with Overlap PCR, and then pTAR-GET-mM-CSF-Δof 30 amino acide located in the intracellular region of brachytmema mutation was obtained; and mean-while, they were identified by PCR and DNA sequencing.The empty vector pTARGET, pTARGET-mM-CSF and pTAR-GET-mM-CSF-Δplasmid were transfected into Ramos cells, the cell line with stable expression was screened by G418 and was detected by RT-PCR and Western blotting.Proliferation and cell cycle of these stably transfected cells were detected by MTT and flow cytometry.Results The mM-CSF and mM-CSF-Δeukaryotic expression vectors were successfully construc-ted.The stable transfectants Ramos-V, Ramos-M and Ramos-Δwere obtained.The OD of proliferation ability of Ramos-M, Ramos-Δand Ramos-V were 0.413 ±0.014, 0.384 ±0.019 and 0.463 ±0.037, respectively.Significant difference was found between the cell lines of Ramos-M and Ramos-Δ, among the cell lines of Ramos-M, Ramos-Δand Ramos-V, re-spectively （P〈0.05 or P 〈0.01）.The proportion of G0/G1 phases of Ramos-M, Ramos-Δand Ramos-V cells were 41.54%±1.22%, 45.60%±1.09% and 39.20% ±1.53%, respectively.Significant difference was found between 〈br〉 Ramos-M and Ramos-Δcells, among Ramos-M, Ramos-Δand Ramos-V cells （P〈0.05 or P〈0.01）.Conclusion mM-CSF and mM-CSF-Δmay inhibit the proliferation of lymphocytic leukemia cell line Ramos and the inhibitory effect of the latter is stronger.