Department of Hematology, Kumamoto University School of Medicine, Kumamoto 860-8556, Japan.
Int J Oncol. 2013 Dec;43(6):1809-16. doi: 10.3892/ijo.2013.2134. Epub 2013 Oct 10.
Although CD138 expression is a hallmark of plasma cells and myeloma cells, reduced CD138 expression is occasionally found. However, the mechanisms underlying CD138 downregulation in myeloma cells remain unclear. Previous reports suggest that the bone marrow microenvironment may contribute to CD138 downregulation. Among various factors in the tumor microenvironment, hypoxia is associated with tumor progression, poor clinical outcomes, dedifferentiation and the formation of cancer stem cell niches in solid tumors. Since recent findings showed that progression of multiple myeloma (MM) delivers hypoxia within the bone marrow, we hypothesized that CD138 expression may be regulated by hypoxia. In the present study, we examined whether the expression of CD138 and transcription factors occurred in myeloma cells under hypoxic conditions. MM cell lines (KMS-12BM and RPMI 8226) were cultured under normoxic or hypoxic conditions for up to 30 days. Changes in the phenotype and the expression of surface antigens and transcription factors were analyzed using flow cytometry, RT-PCR and western blotting. All-trans retinoic acid (ATRA) was used to examine the phenotypic changes under hypoxic conditions. The expression levels of CD138, CS1 and plasma cell-specific transcription factors decreased under hypoxic conditions, while those of CD20, CXCR4 and B cell-specific transcription factors increased compared with those under normoxic conditions. Stem cell-specific transcription factors were upregulated under hypoxic conditions, while no difference was observed in ALDH activity. The reduced CD138 expression under hypoxic conditions recovered when cells were treated with ATRA, even under hypoxic conditions, along with decreases in the expression of stem cell-specific transcription factor. Interestingly, ATRA treatment sensitized MM cells to bortezomib under hypoxia. We propose that hypoxia induces immature and stem cell-like transcription phenotypes in myeloma cells. Taken together with our previous observation that decreased CD138 expression is correlated with disease progression, the present data suggest that a hypoxic microenvironment affects the phenotype of MM cells, which may correlate with disease progression.
虽然 CD138 的表达是浆细胞和骨髓瘤细胞的标志,但偶尔也会发现 CD138 的表达降低。然而,骨髓瘤细胞中 CD138 下调的机制尚不清楚。先前的报告表明,骨髓微环境可能导致 CD138 下调。在肿瘤微环境中的各种因素中,缺氧与肿瘤进展、不良临床结局、去分化以及实体瘤中癌症干细胞龛的形成有关。由于最近的研究结果表明,多发性骨髓瘤(MM)的进展会导致骨髓内缺氧,我们假设 CD138 的表达可能受到缺氧的调节。在本研究中,我们研究了在缺氧条件下骨髓瘤细胞中 CD138 和转录因子的表达情况。将 KMS-12BM 和 RPMI 8226 骨髓瘤细胞系在常氧或缺氧条件下培养长达 30 天。通过流式细胞术、RT-PCR 和 Western blot 分析细胞表型和表面抗原及转录因子的表达变化。使用全反式维甲酸(ATRA)在缺氧条件下研究表型变化。在缺氧条件下,CD138、CS1 和浆细胞特异性转录因子的表达水平降低,而 CD20、CXCR4 和 B 细胞特异性转录因子的表达水平升高。在缺氧条件下,干细胞特异性转录因子上调,而 ALDH 活性无差异。当细胞用 ATRA 处理时,即使在缺氧条件下,CD138 的表达也会恢复,同时干细胞特异性转录因子的表达降低。有趣的是,ATRA 处理在缺氧条件下增加了硼替佐米对 MM 细胞的敏感性。我们提出,缺氧诱导骨髓瘤细胞中不成熟和干细胞样转录表型。结合我们之前的观察结果,即 CD138 表达降低与疾病进展相关,本数据表明,缺氧微环境会影响 MM 细胞的表型,这可能与疾病进展有关。