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Hedgehog Inhibitors in Rhabdomyosarcoma: A Comparison of Four Compounds and Responsiveness of Four Cell Lines.

Ridzewski R, Rettberg D, Dittmann K, Cuvelier N, Fulda S, Hahn H - Front Oncol (2015)

Bottom Line: Our data show that the antitumoral effects of these SMO inhibitors are highly divers and do not necessarily correlate with inhibition of HH signaling.In addition, the effects of SMO inhibitors are concentration-dependent (e.g., 1 and 10 μM GDC-0449 decrease GLI1 expression in RD cells whereas 30 μM GDC-0449 does not).Together these data show that some SMO inhibitors can induce strong antitumoral effects in some, but not all, RMS cell lines.

View Article: PubMed Central - PubMed

Affiliation: Institute of Human Genetics, University Medical Center Goettingen , Goettingen , Germany.

ABSTRACT
Rhabdomyosarcoma (RMS) is the most common soft tissue sarcoma in children and is divided into two major histological subgroups, i.e., embryonal (ERMS) and alveolar RMS (ARMS). RMS can show HEDGEHOG/SMOOTHENED (HH/SMO) signaling activity and several clinical trials using HH inhibitors for therapy of RMS have been launched. We here compared the antitumoral effects of the SMO inhibitors GDC-0449, LDE225, HhA, and cyclopamine in two ERMS (RD, RUCH-2) and two ARMS (RMS-13, Rh41) cell lines. Our data show that the antitumoral effects of these SMO inhibitors are highly divers and do not necessarily correlate with inhibition of HH signaling. In addition, the responsiveness of the RMS cell lines to the drugs is highly heterogeneous. Whereas some SMO inhibitors (i.e., LDE225 and HhA) induce strong proapoptotic and antiproliferative effects in some RMS cell lines, others paradoxically induce cellular proliferation at certain concentrations (e.g., 10 μM GDC-0449 or 5 μM cyclopamine in RUCH-2 and Rh41 cells) or can increase HH signaling activity as judged by GLI1 expression (i.e., LDE225, HhA, and cyclopamine). Similarly, some drugs (e.g., HhA) inhibit PI3K/AKT signaling or induce autophagy (e.g., LDE225) in some cell lines, whereas others cannot (e.g., GDC-0449). In addition, the effects of SMO inhibitors are concentration-dependent (e.g., 1 and 10 μM GDC-0449 decrease GLI1 expression in RD cells whereas 30 μM GDC-0449 does not). Together these data show that some SMO inhibitors can induce strong antitumoral effects in some, but not all, RMS cell lines. Due to the highly heterogeneous response, we propose to conduct thorough pretesting of SMO inhibitors in patient-derived short-term RMS cultures or patient-derived xenograft mouse models before applying these drugs to RMS patients.

No MeSH data available.


Related in: MedlinePlus

BrdU incorporation in RD, RUCH-2, RMS-13, and Rh41 cells after incubation with GDC-0449, LDE225, HhA, and cyclopamine (cp) at the concentration indicated. BrdU-incorporation is shown as percentage of respective solvent-controls (solv) that were set to 100%. The data are represented as mean ± SEM; *p < 0.05; **p < 0.01; ***p < 0.001.
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Figure 2: BrdU incorporation in RD, RUCH-2, RMS-13, and Rh41 cells after incubation with GDC-0449, LDE225, HhA, and cyclopamine (cp) at the concentration indicated. BrdU-incorporation is shown as percentage of respective solvent-controls (solv) that were set to 100%. The data are represented as mean ± SEM; *p < 0.05; **p < 0.01; ***p < 0.001.

Mentions: As shown by BrdU assays, the antiproliferative effects of SMO inhibitors dramatically varied between the cell lines: in RD cells, 30 μM LDE225 and 10 μM and 30 μM HhA reduced the proliferation rate of RD cells approximately by 55% and 30 and 50%, respectively (Figure 2). Thirty micromoles of GDC-0449, 10 μM LDE225 and cyclopamine did not show any antiproliferative effects, whereas 10 μM GDC-0449 significantly increased the proliferation rate (Figure 2). This was similar in RUCH-2 cells, in which the proliferation was significantly reduced by 30 μM LDE225 and by 10 and 30 μM HhA (Figure 2). Ten micromoles of LDE225 and 30 μM GDC-0449 had no effects, whereas 10 μM GDC-0449 and 5 μM cyclopamine significantly increased it (by approximately 50 and 100%, respectively) (Figure 2). In RMS-13, proliferation was reduced by 30 μM GDC-0449, by 10 and 30 μM LDE225 and by 30 μM HhA (Figure 2). In Rh41, none of the drugs inhibited proliferation (Figure 2). However, proliferation-inducing effects were seen for 10 and 30 μM GDC-0449, for 10 μM LDE225 and for cyclopamine (Figure 2; for RD and Rh41 the results were confirmed by simple cell counting; see Figure S5 in Supplementary Material).


Hedgehog Inhibitors in Rhabdomyosarcoma: A Comparison of Four Compounds and Responsiveness of Four Cell Lines.

Ridzewski R, Rettberg D, Dittmann K, Cuvelier N, Fulda S, Hahn H - Front Oncol (2015)

BrdU incorporation in RD, RUCH-2, RMS-13, and Rh41 cells after incubation with GDC-0449, LDE225, HhA, and cyclopamine (cp) at the concentration indicated. BrdU-incorporation is shown as percentage of respective solvent-controls (solv) that were set to 100%. The data are represented as mean ± SEM; *p < 0.05; **p < 0.01; ***p < 0.001.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4459089&req=5

Figure 2: BrdU incorporation in RD, RUCH-2, RMS-13, and Rh41 cells after incubation with GDC-0449, LDE225, HhA, and cyclopamine (cp) at the concentration indicated. BrdU-incorporation is shown as percentage of respective solvent-controls (solv) that were set to 100%. The data are represented as mean ± SEM; *p < 0.05; **p < 0.01; ***p < 0.001.
Mentions: As shown by BrdU assays, the antiproliferative effects of SMO inhibitors dramatically varied between the cell lines: in RD cells, 30 μM LDE225 and 10 μM and 30 μM HhA reduced the proliferation rate of RD cells approximately by 55% and 30 and 50%, respectively (Figure 2). Thirty micromoles of GDC-0449, 10 μM LDE225 and cyclopamine did not show any antiproliferative effects, whereas 10 μM GDC-0449 significantly increased the proliferation rate (Figure 2). This was similar in RUCH-2 cells, in which the proliferation was significantly reduced by 30 μM LDE225 and by 10 and 30 μM HhA (Figure 2). Ten micromoles of LDE225 and 30 μM GDC-0449 had no effects, whereas 10 μM GDC-0449 and 5 μM cyclopamine significantly increased it (by approximately 50 and 100%, respectively) (Figure 2). In RMS-13, proliferation was reduced by 30 μM GDC-0449, by 10 and 30 μM LDE225 and by 30 μM HhA (Figure 2). In Rh41, none of the drugs inhibited proliferation (Figure 2). However, proliferation-inducing effects were seen for 10 and 30 μM GDC-0449, for 10 μM LDE225 and for cyclopamine (Figure 2; for RD and Rh41 the results were confirmed by simple cell counting; see Figure S5 in Supplementary Material).

Bottom Line: Our data show that the antitumoral effects of these SMO inhibitors are highly divers and do not necessarily correlate with inhibition of HH signaling.In addition, the effects of SMO inhibitors are concentration-dependent (e.g., 1 and 10 μM GDC-0449 decrease GLI1 expression in RD cells whereas 30 μM GDC-0449 does not).Together these data show that some SMO inhibitors can induce strong antitumoral effects in some, but not all, RMS cell lines.

View Article: PubMed Central - PubMed

Affiliation: Institute of Human Genetics, University Medical Center Goettingen , Goettingen , Germany.

ABSTRACT
Rhabdomyosarcoma (RMS) is the most common soft tissue sarcoma in children and is divided into two major histological subgroups, i.e., embryonal (ERMS) and alveolar RMS (ARMS). RMS can show HEDGEHOG/SMOOTHENED (HH/SMO) signaling activity and several clinical trials using HH inhibitors for therapy of RMS have been launched. We here compared the antitumoral effects of the SMO inhibitors GDC-0449, LDE225, HhA, and cyclopamine in two ERMS (RD, RUCH-2) and two ARMS (RMS-13, Rh41) cell lines. Our data show that the antitumoral effects of these SMO inhibitors are highly divers and do not necessarily correlate with inhibition of HH signaling. In addition, the responsiveness of the RMS cell lines to the drugs is highly heterogeneous. Whereas some SMO inhibitors (i.e., LDE225 and HhA) induce strong proapoptotic and antiproliferative effects in some RMS cell lines, others paradoxically induce cellular proliferation at certain concentrations (e.g., 10 μM GDC-0449 or 5 μM cyclopamine in RUCH-2 and Rh41 cells) or can increase HH signaling activity as judged by GLI1 expression (i.e., LDE225, HhA, and cyclopamine). Similarly, some drugs (e.g., HhA) inhibit PI3K/AKT signaling or induce autophagy (e.g., LDE225) in some cell lines, whereas others cannot (e.g., GDC-0449). In addition, the effects of SMO inhibitors are concentration-dependent (e.g., 1 and 10 μM GDC-0449 decrease GLI1 expression in RD cells whereas 30 μM GDC-0449 does not). Together these data show that some SMO inhibitors can induce strong antitumoral effects in some, but not all, RMS cell lines. Due to the highly heterogeneous response, we propose to conduct thorough pretesting of SMO inhibitors in patient-derived short-term RMS cultures or patient-derived xenograft mouse models before applying these drugs to RMS patients.

No MeSH data available.


Related in: MedlinePlus