Department of Hematology, XinQiao Hospital, Third Military Medical University, ChongQing, China.
Braz J Med Biol Res. 2012 Oct;45(10):906-12. doi: 10.1590/s0100-879x2012007500107.
Hypoxia inducible factor-1α (HIF-1α) is an important transcription factor, which plays a critical role in the formation of solid tumor and its microenvironment. The objective of the present study was to evaluate the expression and function of HIF-1α in human leukemia bone marrow stromal cells (BMSCs) and to identify the downstream targets of HIF-1α. HIF-1α expression was detected at both the RNA and protein levels using real-time PCR and immunohistochemistry, respectively. Vascular endothelial growth factor (VEGF) and stromal cell-derived factor-1α (SDF-1α) were detected in stromal cells by enzyme-linked immunosorbent assay. HIF-1α was blocked by constructing the lentiviral RNAi vector system and infecting the BMSCs. The Jurkat cell/BMSC co-cultured system was constructed by putting the two cells into the same suitable cultured media and conditions. Cell adhesion and secretion functions of stromal cells were evaluated after transfection with the lentiviral RNAi vector of HIF-1α. Increased HIF-1α mRNA and protein was detected in the nucleus of the acute myeloblastic and acute lymphoblastic leukemia compared with normal BMSCs. The lentiviral RANi vector for HIF-1α was successfully constructed and was applied to block the expression of HIF-1α. When HIF-1α of BMSCs was blocked, the expression of VEGF and SDF-1 secreted by stromal cells were decreased. When HIF-1α was blocked, the co-cultured Jurkat cell's adhesion and migration functions were also decreased. Taken together, these results suggest that HIF-1α acts as an important transcription factor and can significantly affect the secretion and adhesion functions of leukemia BMSCs.
缺氧诱导因子-1α(HIF-1α)是一种重要的转录因子,在实体瘤及其微环境的形成中发挥着关键作用。本研究旨在评估 HIF-1α在人白血病骨髓基质细胞(BMSCs)中的表达和功能,并鉴定其下游靶点。分别采用实时 PCR 和免疫组织化学检测 HIF-1α的 RNA 和蛋白质表达。通过酶联免疫吸附试验检测基质细胞中血管内皮生长因子(VEGF)和基质细胞衍生因子-1α(SDF-1α)的表达。通过构建慢病毒 RNAi 载体系统并感染 BMSCs 来阻断 HIF-1α。将两种细胞置于相同的适宜培养条件下构建 Jurkat 细胞/BMSC 共培养系统。转染 HIF-1α 的慢病毒 RNAi 载体后,评估基质细胞的黏附和分泌功能。与正常 BMSCs 相比,急性髓系白血病和急性淋巴细胞白血病的细胞核中 HIF-1α mRNA 和蛋白表达增加。成功构建了 HIF-1α 的慢病毒 RANi 载体,并应用该载体阻断 HIF-1α 的表达。当 BMSCs 中的 HIF-1α 被阻断时,基质细胞分泌的 VEGF 和 SDF-1 的表达减少。当 HIF-1α 被阻断时,共培养的 Jurkat 细胞的黏附和迁移功能也降低。综上所述,这些结果表明 HIF-1α 作为一种重要的转录因子,可显著影响白血病 BMSCs 的分泌和黏附功能。