Laboratory of Transgenic Models of Diseases, Institute of Molecular Genetics of the Czech Academy of Sciences, Prague, 14220, Czech Republic.
Czech Centre for Phenogenomics Institute of Molecular Genetics of the Czech Academy of Sciences, Prague, 14220, Czech Republic.
Sci Rep. 2019 Apr 19;9(1):6342. doi: 10.1038/s41598-019-42684-y.
Rhabdomyosarcomas (RMS) are a heterogeneous group of mesodermal tumors, the most common sub-types are embryonal (eRMS) and alveolar (aRMS) rhabdomyosarcoma. Immunohistochemical analysis revealed c-Myb expression in both eRMS and aRMS. c-Myb has been reported to be often associated with malignant human cancers. We therefore investigated the c-Myb role in RMS using cellular models of RMS. Specific suppression of c-Myb by a lentiviral vector expressing doxycycline (Dox)-inducible c-Myb shRNA inhibited proliferation, colony formation, and migration of the eRMS cell line (RD), but not of the aRMS cell line (RH30). Upon c-Myb knockdown in eRMS cells, cells accumulated in G0/G1 phase, the invasive behaviour of cells was repressed, and elevated levels of myosin heavy chain, marker of muscle differentiation, was detected. Next, we used an RD-based xenograft model to investigate the role of c-Myb in eRMS tumorigenesis in vivo. We found that Dox administration did not result in efficient suppression of c-Myb in growing tumors. However, when c-Myb-deficient RD cells were implanted into SCID mice, we observed inefficient tumor grafting and attenuation of tumor growth during the initial stages of tumor expansion. The presented study suggests that c-Myb could be a therapeutic target in embryonal rhabdomyosarcoma assuming that its expression is ablated.
横纹肌肉瘤 (RMS) 是一组异质性的中胚层肿瘤,最常见的亚型是胚胎性 (eRMS) 和肺泡性 (aRMS) 横纹肌肉瘤。免疫组织化学分析显示 c-Myb 在 eRMS 和 aRMS 中均有表达。c-Myb 已被报道常与恶性人类癌症有关。因此,我们使用 RMS 的细胞模型研究了 c-Myb 在 RMS 中的作用。通过表达强力霉素 (Dox)-诱导型 c-Myb shRNA 的慢病毒载体特异性抑制 c-Myb,抑制了 eRMS 细胞系 (RD) 的增殖、集落形成和迁移,但不抑制 aRMS 细胞系 (RH30)。在 eRMS 细胞中敲低 c-Myb 后,细胞在 G0/G1 期积累,细胞的侵袭行为受到抑制,并检测到肌球蛋白重链的水平升高,这是肌肉分化的标志物。接下来,我们使用基于 RD 的异种移植模型在体内研究 c-Myb 在 eRMS 肿瘤发生中的作用。我们发现,强力霉素给药并没有导致生长中的肿瘤中 c-Myb 的有效抑制。然而,当将缺乏 c-Myb 的 RD 细胞植入 SCID 小鼠时,我们观察到肿瘤移植效率低下,并且在肿瘤扩张的初始阶段肿瘤生长受到抑制。本研究表明,c-Myb 可能是胚胎性横纹肌肉瘤的治疗靶点,假设其表达被消除。