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目的比较 U937细胞与人骨髓间充质干细胞(MSC)黏附培养前后凋亡相关基因表达谱的变化,寻找白备病耐药与造血微环境的关系。方法 U937细胞分别悬浮培养和与 MSC 黏附培养48h,以流式细胞术测定细胞周期。采用基因表达谱芯片技术测定并分析黏附培养组与悬浮培养组间凋亡基因表达的差异。结果 U937细胞黏附培养与悬浮培养48h,G_0/G_1期细胞分别为(45.3±3.1)%和(32.6±2.1)%(P<0.05),G_2/M 期细胞分别占(2.7±0.3)%和(14.3±1.4)%(P<0.01),自然凋亡率分别为(18.4±1.5)%和(23.4±1.9)%(P<0.05)。在487个凋亡相关候选基因中,筛选出28个明显差异表达基因,上调基因27个,主要分类为凋亡抑制基因、促凋亡基因、细胞周期正调控基因及细胞周期负调控基因等,但以 Bcl-XL 上调最明显。1个下调基因是促凋亡基因。结论与MSC 黏附培养可导致 U937细胞生长抑制,自然凋亡率下降,其机制是多种基因作用的结果,但最主要的可能是 Bcl-XL 凋亡抑制基因上调。
Objective To compare the changes of the expression profiles of apoptosis-related genes between U937 cells and human bone marrow mesenchymal stem cells (MSCs) before and after adhesion, and to find out the relationship between drug resistance and hematopoietic microenvironment. Methods U937 cells were cultured in suspension and adherent culture with MSC for 48h. The cell cycle was determined by flow cytometry. The gene expression profile microarray was used to determine and analyze the difference of apoptosis gene expression between adhesion and suspension culture groups. Results The number of cells in G_0 / G_1 phase of U937 cells was (45.3 ± 3.1)% and (32.6 ± 2.1)% respectively (P <0.05), and cells in G_2 / M phase were (2.7 ± 0.3)% and (14.3 ± 1.4)% (P <0.01). The rates of natural apoptosis were (18.4 ± 1.5)% and (23.4 ± 1.9)%, respectively (P <0.05). Among the 487 apoptosis-related candidate genes, 28 differentially expressed and 27 up-regulated genes were screened, which were mainly classified into apoptosis-inhibitory genes, pro-apoptotic genes, positive cell cycle regulators and cell cycle negative regulatory genes, But the most obvious increase in Bcl-XL. A down-regulated gene is a pro-apoptotic gene. Conclusion Adherent culture with MSCs can induce the growth inhibition of U937 cells and decrease the rate of natural apoptosis. The mechanism may be the result of multiple genes, but the most important one may be the up-regulation of Bcl-XL apoptosis-suppressing genes.