Department of Oncology-Pathology, Karolinska Institutet, Stockholm, Sweden.
Br J Haematol. 2010 Nov;151(4):327-35. doi: 10.1111/j.1365-2141.2010.08362.x. Epub 2010 Aug 31.
We have previously demonstrated that ROR1 and FMOD (fibromodulin) are two genes upregulated in chronic lymphocytic leukaemia (CLL) cells compared to normal blood B cells. In this study, siRNAs were used to specifically silence ROR1 and FMOD expression in CLL cells, healthy B cells and human fibroblast cell lines. siRNA treatment induced a specific reduction (75-95%) in FMOD and ROR1 mRNA. Western blot analysis with specific antibodies for FMOD and ROR1 demonstrated that the proteins were significantly downregulated 48 h after siRNA treatment. Silencing of FMOD and ROR1 resulted in statistically significant (P ≤ 0·05-0·001) apoptosis of CLL cells but not of B cells from normal donors. Human fibroblast cell lines treated with FMOD and ROR1 siRNA did not undergo apoptosis. This is the first report demonstrating that ROR1 and FMOD may be involved in the survival of CLL cells. ROR1 in particular is further explored as potential target for therapy in CLL.
我们之前已经证明，与正常血液 B 细胞相比，ROR1 和 FMOD（纤连蛋白）在慢性淋巴细胞白血病（CLL）细胞中上调。在这项研究中，使用 siRNA 特异性沉默 CLL 细胞、健康 B 细胞和人成纤维细胞系中的 ROR1 和 FMOD 表达。siRNA 处理诱导 FMOD 和 ROR1 mRNA 特异性减少（75-95%）。用针对 FMOD 和 ROR1 的特异性抗体进行 Western blot 分析表明，蛋白质在 siRNA 处理后 48 小时明显下调。沉默 FMOD 和 ROR1 可导致 CLL 细胞但不会导致正常供体的 B 细胞发生统计学上显著的（P ≤ 0·05-0·001）凋亡。用 FMOD 和 ROR1 siRNA 处理的人成纤维细胞系不会发生凋亡。这是第一个表明 ROR1 和 FMOD 可能参与 CLL 细胞存活的报告。ROR1 特别是进一步探索为 CLL 的潜在治疗靶点。
