<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
Although the hydrophobic-polar (HP) model is a simple model to study protein folding, it is an approximation to the real-life case. Dermaseptin is a subfamily of frog skin active peptide family, which has various anti...Although the hydrophobic-polar (HP) model is a simple model to study protein folding, it is an approximation to the real-life case. Dermaseptin is a subfamily of frog skin active peptide family, which has various antimicrobial activities, and dermaseptin-J2 is a newly found peptide composed of 26 amino acids. In this study, the 2-dimensional HP model was used to analyze the foldings of dermaseptin-J2 and its nine mutants, which were converted to different HP sequences according to the normalized amino acid hydrophobicity index with respect to pH levels and the conversion of glycine as hydrophobic or polar, and each has 847,288,609,443 possible foldings. The results show that the foldings with minimal energy have different native states, which are chiral and can be numerically distinguished and ranked according to the normalized amino acid hydrophobicity index. The nine mutants of dermaseptin-J2 do not affect the minimal energy but affect their native states at pH 7. The results demonstrate that two pH levels and conversion of glycine as hydrophobic or polar affect the native state and minimal energy, suggesting these are two ways to modify dermaseptin-J2.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>.
文摘Although the hydrophobic-polar (HP) model is a simple model to study protein folding, it is an approximation to the real-life case. Dermaseptin is a subfamily of frog skin active peptide family, which has various antimicrobial activities, and dermaseptin-J2 is a newly found peptide composed of 26 amino acids. In this study, the 2-dimensional HP model was used to analyze the foldings of dermaseptin-J2 and its nine mutants, which were converted to different HP sequences according to the normalized amino acid hydrophobicity index with respect to pH levels and the conversion of glycine as hydrophobic or polar, and each has 847,288,609,443 possible foldings. The results show that the foldings with minimal energy have different native states, which are chiral and can be numerically distinguished and ranked according to the normalized amino acid hydrophobicity index. The nine mutants of dermaseptin-J2 do not affect the minimal energy but affect their native states at pH 7. The results demonstrate that two pH levels and conversion of glycine as hydrophobic or polar affect the native state and minimal energy, suggesting these are two ways to modify dermaseptin-J2.
