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Updated: Aug 6, 2026
Quantitative PCR-based Assay to Measure Sonic Hedgehog Signaling in Cellular Model of Ciliogenesis
Published on: January 31, 2025
Treatment of eyelid epithelial neoplasm by targeting sonic hedgehog signaling: an experimental study
Ken-Ichi Miyazaki1, Shizuya Saika2, Osamu Yamanaka2
1Department of Ophthalmology, Wakayama Medical University School of Medicine, Wakayama, Japan. miyaken@wakayama-med.ac.jp.
Purpose:
To evaluate the effect of cyclopamine, an inhibitor of the Sonic hedgehog (Shh) signal, on the growth of an epithelial neoplasm.
Methods:
Chemically induced eyelid tumors in XPC-null mice (n=40) were treated daily with a subcutaneous injection of cyclopamine (1 mg/animal) for 7 days. The animals were killed after bromodeoxyuridine (BrdU) labeling, and the tumors were histologically examined. An in vitro study was conducted by using a squamous cell carcinoma (SCC) cell line. The SCC cells were treated with 0, 12.5, or 25.0 microg/ml recombinant Shh (rShh) and either 0 or 100 microM cyclopamine, and cell proliferation was evaluated by using an MTT assay. Cells from this cell line were also implanted subcutaneously in nude mice (n=8) to develop tumors, and the effect of cyclopamine administration was examined in the developed tumors.
Results:
Histology showed that cyclopamine treatment suppressed BrdU incorporation and induced apoptosis in the majority of cells in tumors chemically induced in the eyelid of the XPC-null mice. Cell proliferation of the SCC cell line was enhanced by adding rShh, and this effect was abolished by adding cyclopamine. Proliferation of the SCC cell line was not affected by adding cyclopamine in the absence of rShh. On the other hand, the SCC cells expressed Shh in vivo in tumors developed in nude mice, but cyclopamine suppressed cell proliferation in the tumors, and the Shh-signaling pathway was inhibited by cyclopamine-induced apoptosis.
Conclusions:
Cyclopamine inhibits proliferation and induces apoptosis in epithelial tumor cells in vivo. The Shh-signaling pathway may be a potential therapeutic target for patients with eyelid tumors.
Insights
Cyclopamine, a Sonic hedgehog (Shh) signal inhibitor, effectively suppressed epithelial tumor growth and induced apoptosis in preclinical models. This suggests the Shh pathway is a promising therapeutic target for eyelid tumors.
Area of Science:
- Oncology
- Molecular Biology
- Dermatology
Background:
- Epithelial neoplasms, such as eyelid tumors, represent a significant clinical challenge.
- The Sonic hedgehog (Shh) signaling pathway plays a crucial role in various developmental processes and has been implicated in the pathogenesis of certain cancers.
- Targeting specific signaling pathways offers a promising strategy for novel cancer therapies.
Purpose of the Study:
- To investigate the therapeutic potential of cyclopamine, a Shh signal inhibitor, in controlling epithelial neoplasm growth.
- To elucidate the effects of cyclopamine on cell proliferation and apoptosis in preclinical models of eyelid tumors.
Main Methods:
- In vivo studies involved chemically induced eyelid tumors in XPC-null mice treated with cyclopamine, followed by histological examination and bromodeoxyuridine (BrdU) labeling.
- In vitro experiments utilized a squamous cell carcinoma (SCC) cell line treated with varying concentrations of recombinant Shh (rShh) and cyclopamine, with cell proliferation assessed via MTT assay.
- Tumorigenicity was further evaluated by implanting SCC cells into nude mice and subsequently treating developed tumors with cyclopamine.
Main Results:
- Cyclopamine treatment significantly suppressed BrdU incorporation and induced apoptosis in eyelid tumors in XPC-null mice.
- In vitro, rShh enhanced SCC cell proliferation, an effect abrogated by cyclopamine co-treatment.
- Cyclopamine inhibited proliferation and induced apoptosis in SCC cell line tumors grown in nude mice, demonstrating its efficacy in vivo.
Conclusions:
- Cyclopamine demonstrates potent anti-proliferative and pro-apoptotic effects on epithelial tumor cells in vivo.
- The Shh-signaling pathway is a viable therapeutic target for the management of eyelid tumors.
- These findings support further investigation of cyclopamine as a potential treatment for epithelial neoplasms.
