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Hedgehog signal inhibitors suppress the invasion of human rhabdomyosarcoma cells.
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Two kinds of specific Hh signaling inhibitors, cyclopamine and forskolin, were used to suppress activated Hh signals in three RMS cell lines.The number of invaded cells counted in six random microscopic fields in the Matrigel chambers was significantly decreased by both cyclopamine and forskolin in every RMS cell line.Hh inhibitors may provide a new paradigm for the treatment of RMS.
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PubMed Central - PubMed
Affiliation: Division of Pediatric Surgery, Department of Surgery, Osaka University Graduate School of Medicine, 2-2 Yamadaoka, Suita, Osaka, 565-0871, Japan, ooue@pedsurg.med.osaka-u.ac.jp.
ABSTRACT
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Purpose: In the treatment of rhabdomyosarcoma (RMS), invasion and metastasis remain the most critical determinants of resectability and survival. The objective of this study was to determine whether Hedgehog (Hh) signaling plays a role in the invasion of RMS. Methods: Two kinds of specific Hh signaling inhibitors, cyclopamine and forskolin, were used to suppress activated Hh signals in three RMS cell lines. The effects of the Hh signaling inhibitors on tumor cell invasion and motility were investigated using Matrigel invasion assays and wound closure assays, respectively. Results: The number of invaded cells counted in six random microscopic fields in the Matrigel chambers was significantly decreased by both cyclopamine and forskolin in every RMS cell line. Furthermore, the wound closure assays revealed that a blockade of the Hh signaling pathway by the Hh inhibitors strongly impairs RMS cell motility, as visualized by the delayed closure of the gaps generated in the cultured cell monolayers of the three RMS cell lines. Conclusions: Both the invasive capacity and motility of RMS cells are significantly suppressed by Hh signaling inhibitors, demonstrating that the Hh pathway plays an important role in the invasion of RMS. Hh inhibitors may provide a new paradigm for the treatment of RMS. Related in: MedlinePlus |
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Fig3: Number of invading cells in the Matrigel invasion assays The number of invading cells significantly decreased in both cyclopamine and forskolin-treated groups compared with the control groups in the three RMS cell lines Mentions: We employed cyclopamine and forskolin (specific inhibitors of the Hedgehog pathway) to block the Hh pathway in the RMS cell lines and then assessed the changes in the invasive potential of the cells. The Matrigel invasion assays indicated that RD cells exhibit the strongest invasive potential. As shown in Figs. 2 and 3, the number of invaded cells counted in six random microscopic fields in the Matrigel chamber was significantly decreased by both cyclopamine and forskolin in every RMS cell line. The mean invasiveness for the control, cyclopamine-treated and forskolin-treated RMS-YM cells was 145.2 + 55.5, 27.2 + 7.9 and 43.0 + 16.3 cells (P < 0.01)/6 random microscopic fields, respectively. The mean invasiveness for the control, cyclopamine-treated and forskolin-treated RD cells was 190.7 + 67.2, 77.3 + 29.0 and 131.5 + 22.7 cells (P < 0.05)/6 random microscopic fields, respectively. The mean invasiveness for the control, cyclopamine-treated and forskolin-treated RH30 cells was 104.3 + 14.1, 62.2 + 16.0 and 51.3 + 21.9 cells (P < 0.05)/6 random microscopic fields, respectively.Fig. 2 |
View Article: PubMed Central - PubMed
Affiliation: Division of Pediatric Surgery, Department of Surgery, Osaka University Graduate School of Medicine, 2-2 Yamadaoka, Suita, Osaka, 565-0871, Japan, ooue@pedsurg.med.osaka-u.ac.jp.
Purpose: In the treatment of rhabdomyosarcoma (RMS), invasion and metastasis remain the most critical determinants of resectability and survival. The objective of this study was to determine whether Hedgehog (Hh) signaling plays a role in the invasion of RMS.
Methods: Two kinds of specific Hh signaling inhibitors, cyclopamine and forskolin, were used to suppress activated Hh signals in three RMS cell lines. The effects of the Hh signaling inhibitors on tumor cell invasion and motility were investigated using Matrigel invasion assays and wound closure assays, respectively.
Results: The number of invaded cells counted in six random microscopic fields in the Matrigel chambers was significantly decreased by both cyclopamine and forskolin in every RMS cell line. Furthermore, the wound closure assays revealed that a blockade of the Hh signaling pathway by the Hh inhibitors strongly impairs RMS cell motility, as visualized by the delayed closure of the gaps generated in the cultured cell monolayers of the three RMS cell lines.
Conclusions: Both the invasive capacity and motility of RMS cells are significantly suppressed by Hh signaling inhibitors, demonstrating that the Hh pathway plays an important role in the invasion of RMS. Hh inhibitors may provide a new paradigm for the treatment of RMS.
