Dermaseptin B2’s Anti-Proliferative Activity and down Regulation of Anti-Proliferative, Angiogenic and Metastatic Genes in Rhabdomyosarcoma RD Cells <i>in Vitro</i> — Oak Academic Publishing
Research ArticleOpen AccessGoogle Scholar indexed
Dermaseptin B2’s Anti-Proliferative Activity and down Regulation of Anti-Proliferative, Angiogenic and Metastatic Genes in Rhabdomyosarcoma RD Cells <i>in Vitro</i>
Department of Molecular Biology and Biotechnology, Pan African University Institute for Basic Sciences Technology and Innovation (PAUSTI), Nairobi, Kenya
,
Department of Biochemistry, College of Health Sciences, Jomo Kenyatta University of Agriculture and Technology, Nairobi, Kenya
,
Department of Biochemistry, College of Health Sciences, Jomo Kenyatta University of Agriculture and Technology, Nairobi, Kenya
,
Centre for Public Health Research, Kenya Medical Research Institute, Nairobi, Kenya
,
Department of Biochemistry and Molecular Biology, Faculty of Health Sciences, Busitema University, Mbale, Uganda
,
Department of Biochemistry, College of Health Sciences, University of Nairobi, Nairobi, Kenya
1 Department of Molecular Biology and Biotechnology, Pan African University Institute for Basic Sciences Technology and Innovation (PAUSTI), Nairobi, Kenya
2 Department of Biochemistry, College of Health Sciences, Jomo Kenyatta University of Agriculture and Technology, Nairobi, Kenya
3 Department of Biochemistry, College of Health Sciences, Jomo Kenyatta University of Agriculture and Technology, Nairobi, Kenya
4 Centre for Public Health Research, Kenya Medical Research Institute, Nairobi, Kenya
5 Department of Biochemistry and Molecular Biology, Faculty of Health Sciences, Busitema University, Mbale, Uganda
6 Department of Biochemistry, College of Health Sciences, University of Nairobi, Nairobi, Kenya
Background: 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. Aim: Evaluating the in vitro anti-proliferative activity of Dermaseptin B2 on Rhabdomyosarcoma RD (CCL-136TM) cells and its effect on the expression of MYC, FGFR1, NOTCH1 , and CXCR7 genes involve in processes including proliferation, angiogenesis and metastasis. Methods: 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 MYC, NOTCH1, FGFR1 and CXCR7 , total RNA was extracted from the cells and q-RT-PCR was performed with β-Actin as reference gene. Results: Dermaseptin B2 inhibited the proliferation of RD cells in a time and concentration dependent manner as with IC 50 values of 7.679 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 MYC (fc; 1.5013, 1.5185, 2.4144), CXCR7 (fc; 2.8818, 4.4430, 3.9924), FGFR1 (fc; 2.3515, 2.0809, 2.2543), NOTCH1 (fc; 2.4667, 4.6274, 4.3352) genes for the three-time points respectively. NOTCH1 and CXCR7 showed higher fold changes with respect to β-Actin than MYC and FGFR1 . Conclusion: The results of this study indicate that Dermaseptin B2 is 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 in vivo .
Kommoss, F.K.F., Stichel, D., Mora, J., Esteller, M., et al. (2021) Clinicopathologic and Molecular Analysis of Embryonal Rhabdomyosarcoma of the Genitourinary Tract: Evidence for a Distinct DICER1-Associated Subgroup. Modern Pathology, 34, 1558-1569. https://doi.org/10.1038/s41379-021-00804-y
Kather, J.N., Horner, C., Weis, C.A., et al. (2019) CD163+ Immune Cell Infiltrates and Presence of CD54+ Microvessels Are Prognostic Markers for Patients with Embryonal Rhabdomyosarcoma. Scientific Reports, 9, Article No. 9211. https://doi.org/10.1038/s41598-019-45551-y
Liu, Z., Zhang, X., Lei, H., et al. (2020) CASZ1 Induces Skeletal Muscle and Rhabdomyosarcoma Differentiation through a Feed-Forward Loop with MYOD and MYOG. Nature Communications, 11, Article No. 911. https://doi.org/10.1038/s41467-020-14684-4
Pomella, S., Sreenivas, P., Gryder, B.E., et al. (2021) Interaction between SNAI2 and MYOD Enhances Oncogenesis and Suppresses Differentiation in Fusion Negative Rhabdomyosarcoma. Nature Communications, 12, Article No. 192. https://doi.org/10.1038/s41467-020-20386-8
Walter, D., Satheesha, S., Albrecht, P., et al. (2011) CD133 Positive Embryonal Rhabdomyosarcoma Stem-Like Cell Population Is Enriched in Rhabdospheres. PLoS ONE, 6, e19506. https://doi.org/10.1371/journal.pone.0019506
Hoang, N.T., Acevedo, L.A., Mann, M.J. and Tolani, B. (2018) A Review of Soft-Tissue Sarcomas: Translation of Biological Advances into Treatment Measures. Cancer Management and Research, 10, 1089-1114. https://doi.org/10.2147/CMAR.S159641
Chen, C., Dorado Garcia, H., Scheer, M. and Henssen, A.G. (2019) Current and Future Treatment Strategies for Rhabdomyosarcoma. Frontiers in Oncology, 9, 1458. https://doi.org/10.3389/fonc.2019.01458
Bisogno, G., Jenney, M., Bergeron, C., et al. (2018) Addition of Dose-Intensified Doxorubicin to Standard Chemotherapy for Rhabdomyosarcoma (EpSSG RMS 2005): A Multicentre, Open-Label, Randomised Controlled, Phase 3 Trial. The Lancet Oncology, 19, 1061-1071. https://doi.org/10.1016/S1470-2045(18)30337-1
Amer, K.M., Thomson, J.E., Congiusta, D., et al. (2019) Epidemiology, Incidence, and Survival of Rhabdomyosarcoma Subtypes: SEER and ICES Database Analysis. Journal of Orthopaedic Research, 37, 2226-2230. https://doi.org/10.1002/jor.24387
Rossi, F., Legnini, I., Megiorni, F., et al. (2019) Circ-ZNF609 Regulates G1-S Progression in Rhabdomyosarcoma. Oncogene, 38, 3843-3854. https://doi.org/10.1038/s41388-019-0699-4
Lychou, S.E., Gustafsson, G.G. and Ljungman, G.E. (2016) Higher Rates of Metastatic Disease May Explain the Declining Trend in Swedish Paediatric Rhabdomyosarcoma Survival Rates. Acta Paediatrica, 105, 74-81. https://doi.org/10.1111/apa.13172
Li, Y., Bakke, J., Finkelstein, D., Zeng, H., Wu, J. and Chen, T. (2018) HNRNPH1 Is Required for Rhabdomyosarcoma Cell Growth and Survival. Oncogenesis, 7, Article No. 9. https://doi.org/10.1038/s41389-017-0024-4
Marshall, A.D. and Grosveld, G.C. (2012) Alveolar Rhabdomyosarcoma—The Molecular Drivers of PAX3/7-FOXO1-Induced Tumorigenesis. Skeletal Muscle, 2, Article No. 25. https://doi.org/10.1186/2044-5040-2-25
Hinson, A.R.P., Jones, R., Lisa, L.E., Belyea, B.C., Barr, F.G. and Linardic, C.M. (2013) Human Rhabdomyosarcoma Cell Lines for Rhabdomyosarcoma Research: Utility and Pitfalls. Frontiers in Oncology, 3, 183. https://doi.org/10.3389/fonc.2013.00183
Shern, J.F., Chen, L., Chmielecki, J., et al. (2014) Comprehensive Genomic Analysis of Rhabdomyosarcoma Reveals a Landscape of Alterations Affecting a Common Genetic Axis in Fusion-Positive and Fusion-Negative Tumors. Cancer Discovery, 4, 216-231. https://doi.org/10.1158/2159-8290.CD-13-0639
Comiskey, D.F., Jacob, A.G., Sanford, B.L., et al. (2018) A Novel Mouse Model of Rhabdomyosarcoma Underscores the Dichotomy of MDM2-ALT1 Function in Vivo. Oncogene, 37, 95-106. https://doi.org/10.1038/onc.2017.282
Patel, J.H., Loboda, A.P., Showe, M.K., Showe, L.C. and McMahon, S.B. (2004) Analysis of Genomic Targets Reveals Complex Functions of MYC. Nature Reviews Cancer, 4, 562-568. https://doi.org/10.1038/nrc1393
Dang, C.V. (2012) MYC on the Path to Cancer. Cell, 149, 22-35. https://doi.org/10.1016/j.cell.2012.03.003
Feng, Y.C., Liu, X.Y., Teng, L., et al. (2020) c-Myc Inactivation of p53 through the Pan-Cancer lncRNA MILIP Drives Cancer Pathogenesis. Nature Communications, 11, Article No. 4980. https://doi.org/10.1038/s41467-020-18735-8
Missiaglia, E., Selfe, J., Hamdi, M., et al. (2009) Genomic Imbalances in Rhabdomyosarcoma Cell Lines Affect Expression of Genes Frequently Altered in Primary Tumors: An Approach to Identify Candidate Genes Involved in Tumor Development. Genes, Chromosomes & Cancer, 48, 455-467. https://doi.org/10.1002/gcc.20655
Marampon, F., Ciccarelli, C. and Zani, B.M. (2006) Down-Regulation of c-Myc Following MEK/ERK Inhibition Halts the Expression of Malignant Phenotype in Rhabdomyosarcoma and in Non Muscle-Derived Human Tumors. Molecular Cancer, 5, Article No. 31. https://doi.org/10.1186/1476-4598-5-31
Ilic, N., Utermark, T., Widlund, H.R. and Roberts, T.M. (2011) PI3K-Targeted Therapy Can Be Evaded by Gene Amplification along the MYC-Eukaryotic Translation Initiation Factor 4E (eIF4E) Axis. Proceedings of the National Academy of Sciences of the United States of America, 108, E699-E708. https://doi.org/10.1073/pnas.1108237108
Muellner, M.K., Uras, I.Z., Gapp, B.V., et al. (2011) A Chemical-Genetic Screen Reveals a Mechanism of Resistance to PI3K Inhibitors in Cancer. Nature Chemical Biology, 7, 787-793. https://doi.org/10.1038/nchembio.695
Huang, Z., Lin, S., Long, C., et al. (2018) Notch Signaling Pathway Mediates Doxorubicin-Driven Apoptosis in Cancers. Cancer Management and Research, 10, 1439-1448. https://doi.org/10.2147/CMAR.S160315
Belyea, B.C., Naini, S., Bentley, R.C. and Linardic, C.M. (2011) Inhibition of the Notch-Hey1 Axis Blocks Embryonal Rhabdomyosarcoma Tumorigenesis. Clinical Cancer Research, 17, 7324-7336. https://doi.org/10.1158/1078-0432.CCR-11-1004
Kopan, R. (2002) Notch: A Membrane-Bound Transcription Factor. Journal of Cell Science, 115, 1095-1097. https://doi.org/10.1242/jcs.115.6.1095
Roma, J., Masià, A., Reventós, J., De Toledo, J.S. and Gallego, S. (2010) Notch Pathway Inhibition Significantly Reduces Rhabdomyosarcoma Invasiveness and Mobility in Vitro. Clinical Cancer Research, 17, 505-513. https://doi.org/10.1158/1078-0432.CCR-10-0166
Ignatius, M.S., Hayes, M.N., Lobbardi, R., Chen, E.Y., McCarthy, K.M., et al. (2017) The NOTCH1/SNAIL1/MEF2C Pathway Regulates Growth and Self-Renewal in Embryonal Rhabdomyosarcoma. Cell Reports, 19, 2304-2318. https://doi.org/10.1016/j.celrep.2017.05.061
Roma, J., Almazán-Moga, A., Sánchez de Toledo, J. and Gallego, S. (2012) Notch, Wnt, and Hedgehog Pathways in Rhabdomyosarcoma: From Single Pathways to an Integrated Network. Sarcoma, 2012, Article ID: 695603. https://doi.org/10.1155/2012/695603
Korc, M. and Friesel, R. (2009) The Role of Fibroblast Growth Factors in Tumor Growth. Current Cancer Drug Targets, 9, 639-651. https://doi.org/10.2174/156800909789057006
Zhou, W.Y., Zheng, H., Du, X.L. and Yang, J.L. (2016) Characterization of FGFR Signaling Pathway as Therapeutic Targets for Sarcoma Patients. Cancer Biology & Medicine, 13, 260-268. https://doi.org/10.20892/j.issn.2095-3941.2015.0102
Wesche, J., Haglund, K. and Haugsten, E.M. (2011) Fibroblast Growth Factors and Their Receptors in Cancer. Biochemical Journal, 437, 199-213. https://doi.org/10.1042/BJ20101603
Mouron, S., Manso, L., Caleiras, E., et al. (2021) FGFR1 Amplification or Overexpression and Hormonal Resistance in Luminal Breast Cancer: Rationale for a Triple Blockade of ER, CDK4/6, and FGFR1. Breast Cancer Research, 23, Article No. 21. https://doi.org/10.1186/s13058-021-01398-8
Suyama, K., Shapiro, I., Guttman, M. and Hazan, R.B. (2002) A Signaling Pathway Leading to Metastasis Is Controlled by N-Cadherin and the FGF Receptor. Cancer Cell, 2, 301-314. https://doi.org/10.1016/S1535-6108(02)00150-2
Xian, W., Schwertfeger, K.L., Vargo-Gogola, T. and Rosen, J.M. (2005) Pleiotropic Effects of FGFR1 on Cell Proliferation, Survival, and Migration in a 3D Mammary Epithelial Cell Model. Journal of Cell Biology, 171, 663-673. https://doi.org/10.1083/jcb.200505098
Massabeau, C., Sigal-Zafrani, B., Belin, L., et al. (2012) The Fibroblast Growth Factor Receptor 1 (FGFR1), a Marker of Response to Chemoradiotherapy in Breast Cancer? Breast Cancer Research and Treatment, 134, 259-266. https://doi.org/10.1007/s10549-012-2027-3
Chen, L., Qi, H., Zhang, L., et al. (2018) Effects of FGFR Gene Polymorphisms on Response and Toxicity of Cyclophosphamide-Epirubicin-Docetaxel-Based Chemotherapy in Breast Cancer Patients. BMC Cancer, 18, Article No. 1038. https://doi.org/10.1186/s12885-018-4951-z
Goldstein, M., Meller, I. and Orr-Urtreger, A. (2007) FGFR1 Over-Expression in Primary Rhabdomyosarcoma Tumors Is Associated with Hypomethylation of a 5' CpG Island and Abnormal Expression of the AKT1, NOG, and BMP4 Genes. Genes, Chromosomes & Cancer, 46, 1028-1038. https://doi.org/10.1002/gcc.20489
Zhou, Y., Wu, C., Lu, G., Hu, Z., Chen, Q. and Du, X. (2020) FGF/FGFR Signaling Pathway Involved Resistance in Various Cancer Types. Journal of Cancer, 11, 2000-2007. https://doi.org/10.7150/jca.40531
Wang, C., Chen, W. and Shen, J. (2018) CXCR7 Targeting and Its Major Disease Relevance. Frontiers in Pharmacology, 9, 641. https://doi.org/10.3389/fphar.2018.00641
Zheng, K., Li, H.Y., Su, X.L., et al. (2010) Chemokine Receptor CXCR7 Regulates the Invasion, Angiogenesis and Tumor Growth of Human Hepatocellular Carcinoma Cells. Journal of Experimental & Clinical Cancer Research, 29, Article No. 31. https://doi.org/10.1186/1756-9966-29-31
Ramadan, F., Fahs, A., Ghayad, S.E. and Saab, R. (2020) Signaling Pathways in Rhabdomyosarcoma Invasion and Metastasis. Cancer and Metastasis Reviews, 39, 287-301. https://doi.org/10.1007/s10555-020-09860-3
Grymula, K., Tarnowski, M., Wysoczynski, M., et al. (2010) Overlapping and Distinct Role of CXCR7-SDF-1/ITAC and CXCR4-SDF-1 Axes in Regulating Metastatic Behavior of Human Rhabdomyosarcomas. International Journal of Cancer, 127, 2554-2568. https://doi.org/10.1002/ijc.25245
Würth, R., Bajetto, A., Harrison, J.K., Barbieri, F. and Florio, T. (2014) CXCL12 Modulation of CXCR4 and CXCR7 Activity in Human Glioblastoma Stem-Like Cells and Regulation of the Tumor Microenvironment. Frontiers in Cellular Neuroscience, 8, 144. https://doi.org/10.3389/fncel.2014.00144
Simpson, G., Relph, K., Harrington, K., Melcher, A. and Pandha, H. (2016) Cancer Immunotherapy via Combining Oncolytic Virotherapy with Chemotherapy: Recent Advances. Oncolytic Virotherapy, 5, 1-13. https://doi.org/10.2147/OV.S66083
Steckbeck, J.D., Deslouches, B. and Montelaro, R.C. (2014) Antimicrobial Peptides: New Drugs for Bad Bugs? Expert Opinion on Biological Therapy, 14, 11-14. https://doi.org/10.1517/14712598.2013.844227
Deslouches, B., Steckbeck, J.D., Craigo, J.K., Doi, Y., Burns, J.L. and Montelaro, R.C. (2015) Engineered Cationic Antimicrobial Peptides to Overcome Multidrug Resistance by ESKAPE Pathogens. Antimicrobial Agents and Chemotherapy, 59, 1329-1333. https://doi.org/10.1128/AAC.03937-14
Deslouches, B., Steckbeck, J.D., Craigo, J.K., Doi, Y., Mietzner, T.A. and Montelaro, R.C. (2013) Rational Design of Engineered Cationic Antimicrobial Peptides Consisting Exclusively of Arginine and Tryptophan, and Their Activity against Multidrug-Resistant Pathogens. Antimicrobial Agents and Chemotherapy, 57, 2511-2521. https://doi.org/10.1128/AAC.02218-12
Huang, L., Chen, D., Wang, L., et al. (2017) Dermaseptin-PH: A Novel Peptide with Antimicrobial and Anticancer Activities from the Skin Secretion of the South American Orange-Legged Leaf Frog, Pithecopus (Phyllomedusa) Hypochondrialis. Molecules, 22, 1805. https://doi.org/10.3390/molecules22101805
Mangoni, M.L., Papo, N., Saugar, J.M., et al. (2006) Effect of Natural L- to D-Amino Acid Conversion on the Organization, Membrane Binding, and Biological Function of the Antimicrobial Peptides Bombinins H. Biochemistry, 45, 4266-4276. https://doi.org/10.1021/bi052150y
Deslouches, B. and Peter Di, Y. (2017) Antimicrobial Peptides with Selective Antitumor Mechanisms: Prospect for Anticancer Applications. Oncotarget, 8, 46635-46651.
Papo, N., Shahar, M., Eisenbach, L. and Shai, Y. (2003) A Novel Lytic Peptide Composed of DL-Amino Acids Selectively Kills Cancer Cells in Culture and in Mice. Journal of Biological Chemistry, 278, 21018-21023. https://doi.org/10.1074/jbc.M211204200
Papo, N. and Shai, Y. (2003) New Lytic Peptides Based on the D,L-Amphipathic Helix Motif Preferentially Kill Tumor Cells Compared to Normal Cells. Biochemistry, 42, 9346-9354. https://doi.org/10.1021/bi027212o
Ellerby, H., Arap, W., Ellerby, L.M., et al. (1999) Anticancer Activity of Targeted Proapoptotic Peptides. Nature Medicine, 5, 1032-1038. https://doi.org/10.1038/12469
Chen, Y., Xu, X., Hong, S., et al. (2001) RGD-Tachyplesin Inhibits Tumor Growth. Cancer Research, 61, 2434-2438.
Papo, N., Braunstein, A., Eshhar, Z. and Shai, Y. (2004) Suppression of Human Prostate Tumor Growth in Mice by a Cytolytic D-, L-Amino Acid Peptide: Membrane Lysis, Increased Necrosis, and Inhibition of Prostate-Specific Antigen Secretion. Cancer Research, 64, 5779-5786. https://doi.org/10.1158/0008-5472.CAN-04-1438
Papo, N., Seger, D., Makovitzki, A., et al. (2006) Inhibition of Tumor Growth and Elimination of Multiple Metastases in Human Prostate and Breast Xenografts by Systemic Inoculation of a Host Defense-Like Lytic Peptide. Cancer Research, 66, 5371-5378. https://doi.org/10.1158/0008-5472.CAN-05-4569
Daly, J.W., Caceres, J., Moni, R.W., et al. (1992) Frog Secretions and Hunting Magic in the Upper Amazon: Identification of a Peptide That Interacts with an Adenosine Receptor. Proceedings of the National Academy of Sciences of the United States of America, 89, 10960-10963. https://doi.org/10.1073/pnas.89.22.10960
Amiche, M., Ducancel, F., Mor, A., Boulain, J.C., Menez, A. and Nicolas, P. (1994) Precursors of Vertebrate Peptide Antibiotics Dermaseptin b and Adenoregulin Have Extensive Sequence Identities with Precursors of Opioid Peptides Dermorphin, Dermenkephalin, and Deltorphins. Journal of Biological Chemistry, 269, 17847-17852. https://doi.org/10.1016/S0021-9258(17)32386-4
Charpentier, S., Amiche, M., Mester, J., et al. (1998) Structure, Synthesis, and Molecular Cloning of Dermaseptins B, a Family of Skin Peptide Antibiotics. Journal of Biological Chemistry, 273, 14690-14697. https://doi.org/10.1074/jbc.273.24.14690
Van Zoggel, H., Hamma-Kourbali, Y., Galanth, C., et al. (2012) Antitumor and Angiostatic Peptides from Frog Skin Secretions. Amino Acids, 42, 385-395. https://doi.org/10.1007/s00726-010-0815-9
van Zoggel, H., Carpentier, G., Dos Santos, C., et al. (2012) Antitumor and Angiostatic Activities of the Antimicrobial Peptide Dermaseptin B2. PLoS ONE, 7, e44351. https://doi.org/10.1371/journal.pone.0044351
Opanda, S.M., Wamunyokoli, F., Khamadi, S., Coldren, R. and Bulimo, W.D. (2016) Genotyping of Enteroviruses Isolated in Kenya from Pediatric Patients Using Partial VP1 Region. SpringerPlus, 5, Article No. 158. https://doi.org/10.1186/s40064-016-1834-0
O’Brien, J., Wilson, I., Orton, T. and Pognan, F. (2000) Investigation of the Alamar Blue (Resazurin) Fluorescent Dye for the Assessment of Mammalian Cell Cytotoxicity. European Journal of Biochemistry, 267, 5421-5426. https://doi.org/10.1046/j.1432-1327.2000.01606.x
Rodríguez-Corrales, J. and Josan, J.S. (2017) Resazurin Live Cell Assay: Setup and Fine-Tuning for Reliable Cytotoxicity Results. In: Lazar, I., Kontoyianni, M. and Lazar, A., Eds., Methods in Molecular Biology, Vol. 1647, Humana Press, New York, 207-219. https://doi.org/10.1007/978-1-4939-7201-2_14
Gavamukulya, Y., Maina, E.N., El-Shemy, H.A., et al. (2021) Annona muricata Silver Nanoparticles Exhibit Strong Anticancer Activities against Cervical and Prostate Adenocarcinomas through Regulation of CASP9 and the CXCL1/CXCR2 Genes Axis. Tumor Biology, 43, 37-55. https://doi.org/10.3233/TUB-200058
Abdelrahim, M., Baker, C.H., Abbruzzese, J.L., et al. (2007) Regulation of Vascular Endothelial Growth Factor Receptor-1 Expression by Specificity Proteins 1, 3, and 4 in Pancreatic Cancer Cells. Cancer Research, 67, 3286-3294. https://doi.org/10.1158/0008-5472.CAN-06-3831
Justus, C.R., Leffler, N., Ruiz-Echevarria, M. and Yang, L.V. (2014) In Vitro Cell Migration and Invasion Assays. Journal of Visualized Experiments, 88, e51046. https://doi.org/10.3791/51046
Liang, C.C., Park, A.Y. and Guan, J.L. (2007) In Vitro Scratch Assay: A Convenient and Inexpensive Method for Analysis of Cell Migration in Vitro. Nature Protocols, 2, 329-333. https://doi.org/10.1038/nprot.2007.30
Livak, K.J. and Schmittgen, T.D. (2001) Analysis of Relative Gene Expression Data Using Real-Time Quantitative PCR and the 2 -ΔΔC T Method. Methods, 25, 402-408. https://doi.org/10.1006/meth.2001.1262
Conlon, J.M., Woodhams, D.C., Raza, H., et al. (2007) Peptides with Differential Cytolytic Activity from Skin Secretions of the Lemur Leaf Frog Hylomantis lemur (Hylidae: Phyllomedusinae). Toxicon, 50, 498-506. https://doi.org/10.1016/j.toxicon.2007.04.017
Kramer, N., Walzl, A., Unger, C., et al. (2013) In Vitro Cell Migration and Invasion Assays. Mutation Research/Reviews in Mutation Research, 752, 10-24. https://doi.org/10.1016/j.mrrev.2012.08.001
Chen, H., Liu, H. and Qing, G. (2018) Targeting Oncogenic Myc as a Strategy for Cancer Treatment. Signal Transduction and Targeted Therapy, 3, Article No. 5. https://doi.org/10.1038/s41392-018-0008-7
Polakova, K., Polakova, K., Pizova, K., et al. (2015) In Vitro Cytotoxicity Analysis of Doxorubicin-Loaded/Superparamagnetic Iron Oxide Colloidal Nanoassemblies on MCF7 and NIH3T3 Cell Lines. International Journal of Nanomedicine, 10, 949-961. https://doi.org/10.2147/IJN.S72590
Katoh, M. (2016) FGFR Inhibitors: Effects on Cancer Cells, Tumor Microenvironment and Whole-Body Homeostasis (Review). International Journal of Molecular Medicine, 38, 3-15. https://doi.org/10.3892/ijmm.2016.2620
Libura, J., Drukala, J., Majka, M., et al. (2002) CXCR4-SDF-1 Signaling Is Active in Rhabdomyosarcoma Cells and Regulates Locomotion, Chemotaxis, and Adhesion. Blood, 100, 2597-2606. https://doi.org/10.1182/blood-2002-01-0031
Kucia, M., Reca, R., Miekus, K., et al. (2005) Trafficking of Normal Stem Cells and Metastasis of Cancer Stem Cells Involve Similar Mechanisms: Pivotal Role of the SDF-1-CXCR4 Axis. Stem Cells, 23, 879-894. https://doi.org/10.1634/stemcells.2004-0342
Jankowski, K., Kucia, M., Wysoczynski, M., et al. (2003) Both Hepatocyte Growth Factor (HGF) and Stromal-Derived Factor-1 Regulate the Metastatic Behavior of Human Rhabdomyosarcoma Cells, but only HGF Enhances Their Resistance to Radiochemotherapy. Cancer Research, 63, 7926-7935.