<b><span style="font-family:"">Background: </span></b><span style="font-family:"">Rhabdomyosarcoma (RMS) is the most prevalent soft tissue sarcoma in childre...<b><span style="font-family:"">Background: </span></b><span style="font-family:"">Rhabdomyosarcoma (RMS) is the most prevalent soft tissue sarcoma in children, representing approximately 50% of pediatric sarcomas and can develop in any part of the body though more frequently at the extremities. <b>Aim: </b>Evaluating the<i> in vitro</i> anti-proliferative activity of Dermaseptin B2 on Rhabdomyosarcoma RD (CCL-136TM) cells and its effect on the <span>expression of <i>MYC, FGFR1, NOTCH1</i>, and CXCR7 genes involve in processes </span>including proliferation, angiogenesis and metastasis. <b>Methods: </b>RD cells were grown in Dulbecco’s Modified Eagle’s Medium supplemented with 10% Fetal Bovine Serum. Exponentially growing cells were treated with Dermaseptin B2 and Antiproliferative activity was assayed using the resazurin and migration assays at three time-points. In order to determine the gene expression profiles of <i>MYC, NOTCH1, FGFR1 </i>and<i> CXCR7</i>, total RNA was extracted from the cells and q-RT-PCR was performed with <i>β-Actin</i> as reference gene. <b>Results: </b>Dermaseptin B2 inhibited the proliferation of RD cells in a time and concentration dependent manner as with IC<sub>50</sub> values of 7.679</span><span style="font-family:""> </span><span style="font-family:"">μM, 7.235 μM, 5.993 μM. The 2-dimentional wound healing assay showed inhibition of migration and motility of the RD cells at time-points of 6, 24, 48 and 72-hours with the greatest inhibition observed at 72-hours. Dermaseptin B2 downregulated the target <i>MYC</i> (fc;1.5013, 1.5185, 2.4144), <i>CXCR7</i> (fc;2.8818, 4.4430, 3.9924), <i>FGFR1</i> (fc;2.3515, 2.0809, 2.2543), <i>NOTCH1</i> (fc;2.4667, 4.6274, 4.3352) genes for the three-time points respectively. <i>NOTCH1</i> and <i>CXCR7</i> showed higher fold changes with respect to <i>β-Actin</i> than <i>MYC</i> and <i>FGFR1</i>. <b>Conclusion: </b>The results of this study indicate that Dermaseptin B2 </span><span style="font-family:"">is</span><span style="font-family:""> a target molecule for signaling pathways including PI3K/AKT, RTK and NOTCH pathways that could affect the transcription of these genes and overall inhibition of cancer progression. Further studies are needed to give a better understanding of the detailed mechanisms of action as well as the effects of the Dermaseptin B2 peptide <i>in vivo</i>.展开更多
文摘<b><span style="font-family:"">Background: </span></b><span style="font-family:"">Rhabdomyosarcoma (RMS) is the most prevalent soft tissue sarcoma in children, representing approximately 50% of pediatric sarcomas and can develop in any part of the body though more frequently at the extremities. <b>Aim: </b>Evaluating the<i> in vitro</i> anti-proliferative activity of Dermaseptin B2 on Rhabdomyosarcoma RD (CCL-136TM) cells and its effect on the <span>expression of <i>MYC, FGFR1, NOTCH1</i>, and CXCR7 genes involve in processes </span>including proliferation, angiogenesis and metastasis. <b>Methods: </b>RD cells were grown in Dulbecco’s Modified Eagle’s Medium supplemented with 10% Fetal Bovine Serum. Exponentially growing cells were treated with Dermaseptin B2 and Antiproliferative activity was assayed using the resazurin and migration assays at three time-points. In order to determine the gene expression profiles of <i>MYC, NOTCH1, FGFR1 </i>and<i> CXCR7</i>, total RNA was extracted from the cells and q-RT-PCR was performed with <i>β-Actin</i> as reference gene. <b>Results: </b>Dermaseptin B2 inhibited the proliferation of RD cells in a time and concentration dependent manner as with IC<sub>50</sub> values of 7.679</span><span style="font-family:""> </span><span style="font-family:"">μM, 7.235 μM, 5.993 μM. The 2-dimentional wound healing assay showed inhibition of migration and motility of the RD cells at time-points of 6, 24, 48 and 72-hours with the greatest inhibition observed at 72-hours. Dermaseptin B2 downregulated the target <i>MYC</i> (fc;1.5013, 1.5185, 2.4144), <i>CXCR7</i> (fc;2.8818, 4.4430, 3.9924), <i>FGFR1</i> (fc;2.3515, 2.0809, 2.2543), <i>NOTCH1</i> (fc;2.4667, 4.6274, 4.3352) genes for the three-time points respectively. <i>NOTCH1</i> and <i>CXCR7</i> showed higher fold changes with respect to <i>β-Actin</i> than <i>MYC</i> and <i>FGFR1</i>. <b>Conclusion: </b>The results of this study indicate that Dermaseptin B2 </span><span style="font-family:"">is</span><span style="font-family:""> a target molecule for signaling pathways including PI3K/AKT, RTK and NOTCH pathways that could affect the transcription of these genes and overall inhibition of cancer progression. Further studies are needed to give a better understanding of the detailed mechanisms of action as well as the effects of the Dermaseptin B2 peptide <i>in vivo</i>.