Smoothened as a new therapeutic target for human osteosarcoma.
Smoothened as a new therapeutic target for human osteosarcoma.
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DOI：
10.1186/1476-4598-9-5
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发表时间：
2010-01-12
期刊：
影响因子：
37.3
通讯作者：
Komiya S
中科院分区：
文献类型：
--
作者：
Hirotsu M;Setoguchi T;Sasaki H;Matsunoshita Y;Gao H;Nagao H;Kunigou O;Komiya S
Hedgehog信号通路在胚胎发育中起组织者的作用。最近的研究表明，在各种类型的恶性肿瘤中Hedgehog通路的组成性激活。然而，Hedgehog通路如何参与骨肉瘤的发病机制仍不清楚。为了探讨异常Hedgehog通路在骨肉瘤发病机制中的作用，我们研究了Hedgehog通路在骨肉瘤中的表达和激活，并检测了SMOOTHENED（SMO）抑制剂的作用。为了评估Hedgehog通路基因的表达，我们使用骨肉瘤细胞系和骨肉瘤活检标本进行实时PCR和免疫组化。为了评价SMO抑制的效果，我们进行了体外细胞活力、集落形成、细胞周期和体内异种移植模型。Real-time PCR检测结果显示，骨肉瘤细胞系中Sonic hedgehog、Indian hedgehog、PTCH 1、SMO和GLI均过表达。实时荧光定量PCR显示SMO、PTCH 1和GLI 2在骨肉瘤活检标本中过表达。这些发现表明，刺猬通路在骨肉瘤中被激活。SMO特异性抑制剂环巴胺对SMO的抑制可使骨肉瘤的体外生长减慢。细胞周期分析显示，环巴胺促进G1期阻滞。环巴胺降低细胞周期加速因子包括细胞周期蛋白D1、细胞周期蛋白E1、SKP 2和pRb的表达。环巴胺处理后p21 cip 1 w蛋白表达上调。此外，通过SMO shRNA敲低SMO可在体外和体内阻止骨肉瘤生长。这些结果表明，灭活SMO可能是一个有用的方法来治疗骨肉瘤患者。
The Hedgehog signaling pathway functions as an organizer in embryonic development. Recent studies have demonstrated constitutive activation of Hedgehog pathway in various types of malignancies. However, it remains unclear how Hedgehog pathway is involved in the pathogenesis of osteosarcoma. To explore the involvement of aberrant Hedgehog pathway in the pathogenesis of osteosarcoma, we investigated the expression and activation of Hedgehog pathway in osteosarcoma and examined the effect of SMOOTHENED (SMO) inhibition. To evaluate the expression of genes of Hedgehog pathway, we performed real-time PCR and immunohistochemistry using osteosarcoma cell lines and osteosarcoma biopsy specimens. To evaluate the effect of SMO inhibition, we did cell viability, colony formation, cell cycle in vitro and xenograft model in vivo. Real-time PCR revealed that osteosarcoma cell lines over-expressed Sonic hedgehog, Indian hedgehog, PTCH1, SMO, and GLI. Real-time PCR revealed over-expression of SMO, PTCH1, and GLI2 in osteosarcoma biopsy specimens. These findings showed that Hedgehog pathway is activated in osteosarcomas. Inhibition of SMO by cyclopamine, a specific inhibitor of SMO, slowed the growth of osteosarcoma in vitro. Cell cycle analysis revealed that cyclopamine promoted G1 arrest. Cyclopamine reduced the expression of accelerators of the cell cycle including cyclin D1, cyclin E1, SKP2, and pRb. On the other hand, p21cip1 wprotein was up-regulated by cyclopamine treatment. In addition, knockdown of SMO by SMO shRNA prevents osteosarcoma growth in vitro and in vivo. These findings suggest that inactivation of SMO may be a useful approach to the treatment of patients with osteosarcoma.
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影响因子：
82.9
作者：
Dierks, Christine;Grbic, Jovana;Warmuth, Markus
通讯作者：
Warmuth, Markus
影响因子：
5.2
作者：
Fujii, Hiromasa;Honoki, Kanya;Takakura, Yoshinori
通讯作者：
Takakura, Yoshinori
影响因子：
5.7
作者：
Hegde, Ganapati V.;Munger, Corey M.;Joshi, Shantaram S.
通讯作者：
Joshi, Shantaram S.
影响因子：
4
作者：
Iwaya, K;Ogawa, H;Mukai, K
通讯作者：
Mukai, K
影响因子：
16.2
作者：
Hynes, M;Stone, DM;Rosenthal, A
通讯作者：
Rosenthal, A