Department of Orthopaedic Surgery, Graduate School of Medical and Dental Sciences, Kagoshima University, 8-35-1 Sakuragaoka, Kagoshima 890-8520, Japan.
Int J Oncol. 2011 Oct;39(4):899-906. doi: 10.3892/ijo.2011.1076. Epub 2011 Jun 14.
The Hedgehog pathway functions as an organizer in embryonic development. Recent studies have shown that mutation of the PTCH1 gene involved in the Hedgehog pathway affects rhabdomyosarcoma development. However, the expression of Hedgehog pathway molecules in human rhabdomyosarcoma cells has not been well clarified. In addition, the effect of pharmacological inhibition of the Hedgehog pathway is not known. We investigated the expression of the genes involved in the Hedgehog pathway using human rhabdomyosarcoma cell lines and biopsy specimens. Further, we evaluated the effect of pharmacological inhibition of the Hedgehog pathway using cyclopamine or GANT61 by WST assay, cell proliferation assay and cell death detection assay. Real-time PCR revealed that human rhabdomyosarcoma cell lines and biopsy specimens overexpressed the following genes: Sonic hedgehog, Indian hedgehog, Desert hedgehog, PTCH1, SMO, GLI1, GLI2 and ULK3. Immunohistochemistry revealed that rhabdomyosarcoma cell lines and biopsy specimens expressed SMO and GLI2. Inhibition of SMO by cyclopamine slowed the growth of human rhabdomyosarcoma cell lines. Similarly, inhibition of GLI by GANT61 slowed the growth of human rhabdomyosarcoma cell lines. Inhibition of cell proliferation and apoptotic cell death together prevented the growth of rhabdomyosarcoma cells by cyclopamine and GANT61 treatment. Our findings suggest that pharmacological inhibition of the Hedgehog pathway may be a useful approach for treating rhabdomyosarcoma patients.
Hedgehog 通路在胚胎发育中起组织者的作用。最近的研究表明，参与 Hedgehog 通路的 PTCH1 基因突变会影响横纹肌肉瘤的发展。然而，Hedgehog 通路分子在人横纹肌肉瘤细胞中的表达尚未得到很好的阐明。此外，Hedgehog 通路的药理抑制作用尚不清楚。我们使用人横纹肌肉瘤细胞系和活检标本研究了 Hedgehog 通路相关基因的表达。此外，我们通过 WST 测定、细胞增殖测定和细胞死亡检测测定评估了用环巴胺或 GANT61 进行 Hedgehog 通路的药理抑制的效果。实时 PCR 显示，人横纹肌肉瘤细胞系和活检标本过表达以下基因：Sonic hedgehog、Indian hedgehog、Desert hedgehog、PTCH1、SMO、GLI1、GLI2 和 ULK3。免疫组织化学显示横纹肌肉瘤细胞系和活检标本表达 SMO 和 GLI2。SMO 的抑制cyclopamine 减缓了人横纹肌肉瘤细胞系的生长。同样，GLI 的抑制 GANT61 也减缓了人横纹肌肉瘤细胞系的生长。抑制细胞增殖和凋亡性细胞死亡共同阻止了 cyclopamine 和 GANT61 处理对横纹肌肉瘤细胞的生长。我们的研究结果表明，Hedgehog 通路的药理抑制可能是治疗横纹肌肉瘤患者的一种有用方法。
