Daneshmanesh Amir H, Mikaelsson Eva, Jeddi-Tehrani Mahmood, Bayat Ali Ahmad, Ghods Roya, Ostadkarampour Mahyar, Akhondi Mehdi, Lagercrantz Svetlana, Larsson Catharina, Osterborg Anders, Shokri Fazel, Mellstedt Håkan, Rabbani Hodjattallah
Immune and Gene Therapy Lab, CCK, Karolinska Institutet and Karolinska University Hospital Solna, Stockholm, Sweden.
Int J Cancer. 2008 Sep 1;123(5):1190-5. doi: 10.1002/ijc.23587.
Gene profiling studies of patients with chronic lymphocytic leukemia (CLL) has revealed increased expression of Ror1, a cell surface receptor tyrosine kinase. The aim of present study was to analyze gene and protein expression of Ror1 in CLL cells and normal blood leukocytes. Gene expression analysis reverse transcription-polymerase chain reaction of ROR1 revealed that all patients with CLL (n = 100) spontaneously expressed ROR1 mRNA whereas enriched blood B and T cells as well as granulocytes from healthy donors (n = 10) were negative. A strong nonphysiological activation signal (PMA/ionomycin) was required to induce expression in vitro in normal lymphocytes. Major genomic aberrations (mutations or truncation) of ROR1 were not observed. Protein expression was analyzed by Western blot using a panel of polyclonal anti-Ror antibodies as well as flow cytometry. Blood lymphocytes from 18/18 CLL patients, but none of the 10 healthy donors, expressed surface Ror1. The majority of CLL cells exhibited Ror1 surface expression (71% mean; range 36-92%) with a mean fluorescence intensity (MFI) of 20 (range 10-45). The corresponding MFI of CD19 on CLL cells was 26 (range 9-48). There was no difference in the Ror1 protein expression comparing IgVH mutated and unmutated cases as well as progressive and nonprogressive CLL patients. Two different variants of the Ror1 protein, 105 and 130 kDa, were identified. The Ror1 protein expression in patients with CLL but not in normal leukocytes merits further studies of its role in the pathobiology of CLL, which may provide a basis for development of Ror1 directed targeted therapy.
慢性淋巴细胞白血病（CLL）患者的基因谱研究显示，细胞表面受体酪氨酸激酶Ror1的表达增加。本研究的目的是分析Ror1在CLL细胞和正常血液白细胞中的基因和蛋白表达。ROR1的基因表达分析采用逆转录-聚合酶链反应，结果显示所有CLL患者（n = 100）均自发表达ROR1 mRNA，而健康供者的富集血液B细胞、T细胞以及粒细胞（n = 10）均为阴性。在正常淋巴细胞中，需要强烈的非生理性激活信号（佛波酯/离子霉素）才能在体外诱导其表达。未观察到ROR1的主要基因组畸变（突变或截短）。采用一组多克隆抗Ror抗体通过蛋白质印迹法以及流式细胞术分析蛋白表达。18/18例CLL患者的血液淋巴细胞表达表面Ror1，而10例健康供者均未表达。大多数CLL细胞表现出Ror1表面表达（平均71%；范围36 - 92%），平均荧光强度（MFI）为20（范围10 - 45）。CLL细胞上CD19的相应MFI为26（范围9 - 48）。比较IgVH突变和未突变病例以及进展性和非进展性CLL患者，Ror1蛋白表达无差异。鉴定出Ror1蛋白的两种不同变体，分子量分别为105 kDa和130 kDa。CLL患者而非正常白细胞中Ror1蛋白的表达值得进一步研究其在CLL病理生物学中的作用，这可能为开发针对Ror1的靶向治疗提供基础。
