摘要
背景:既往多项研究提示多发性骨髓瘤患者间充质干细胞的细胞因子分泌、免疫调节、成骨分化等生物功能异常。目的:观察白细胞介素1β单次或多次预处理的正常间充质干细胞后,其再与多发性骨髓瘤细胞株共培养,对骨髓瘤细胞干细胞相关基因、趋化相关受体等表达的影响。方法:实验分为无白细胞介素1β预处理的骨髓间充质干细胞对照组、白细胞介素1β预刺激1 d及白细胞介素1β预刺激7 d组,各组分别培养7 d后,均加入骨髓瘤细胞株H929直接接触共培养。3 d后收集各组中骨髓瘤细胞株H929细胞,分别采用Western blot方法检测各组细胞NANOG、SOX2和OCT-4蛋白的表达水平,RT-PCR方法检测各组细胞趋化因子受体CCR1、CCR2、CCR6和CXCR4的基因表达水平。结果与结论:(1)与对照组比较,H929细胞株和单次白细胞介素1β处理组及多次白细胞介素1β处理组共培养后,SOX2蛋白表达水平升高(P<0.05);(2)与对照组、单次白细胞介素1β处理组比较,多次白细胞介素1β处理组CCR1和CCR2基因的表达水平均增高(P<0.05);(3)实验结果显示,正常骨髓间充质干细胞经炎症因子白细胞介素1β单次或多次刺激后,在与骨髓瘤细胞的相互作用中,对骨髓瘤细胞的干细胞基因表达、细胞趋化因子受体表达等均产生影响。这提示骨髓微环境中异常水平炎性因子,除了直接作用于骨髓瘤细胞之外,很可能通过影响间充质干细胞等非骨髓瘤细胞后,间接参与骨髓瘤疾病的发生和发展,并且其对骨髓瘤细胞的影响效果与白细胞介素1β作用间充质干细胞的时间长短有关。
BACKGROUND： Many studies have showed that mesenchymal stem cells derived from multiple myeloma patients display a wide range of abnormal biological features, including cytokine secretion, immune regulation and osteogenic differentiation. OBJECTIVE： To investigate the expression of stem cell-associated proteins and chemotaxis-related receptors genes in myeloma cells when co-cultured with pulsed or sustained interleukin-1β treated bone marrow mesenchymal stem cells. METHODS： There were three groups in the experiment： untreated normal bone marrow mesenchymal stem cells as control, bone marrow mesenchymal stem cells only treated with interleukin-1 β for 1 day as 1-day group or once a day for 7 days as 7-day group. After 7 days of culture, treated or untreated bone marrow mesenchymal stem cells were employed as coating feeder for the direct co-culture with H929 cells for 3 days. At the end of co-culture, H929 myeloma cells were harvested, the expression of NANOG, SOX2 and OCT-4 was measured by western blot, and mRNA levels of CCR1, CCR2, CCR6 and CXCR4 were detected by real-time PCR. RESULTS AND CONCLUSION： The SOX2 protein levels in H929 cells were significantly higher in both 1-day and 7-day groups than that of control group （P 〈 0.05）. In the respective of chemotaxis-related receptors, the mRNA levels of CCR1 and CCR2 in myeloma cells in 7-day group were significantly higher than those in both control and 1-day groups （P 〈 0.05）. Our in vitro results impose inflammatory cytokine interleukin-1β in bone marrow microenvironment has direct effect on myeloma cells but also regulates rnyeloma cells indirectly via non-myeloma cells such as mesenchymal stern cells; besides, sustained elevated intedeukin-lβ level in the bone marrow has more profound effects on myeloma cells through mesenchymal stem cells. Mesenchymal stem cells act an important cellular component involved in myeloma progression.
出处
《中国组织工程研究》
CAS
北大核心
2017年第1期54-59,共6页
Chinese Journal of Tissue Engineering Research
基金
国家自然科学基金(81202358
81571582)
江苏省卫生计生科研项目(Z201512)~~
