Small interfering RNA‐mediated CXCR1 or CXCR2 knock‐down inhibits melanoma tumor growth and invasion

• Published in: International journal of cancer 2010-01, Vol.126 (2), p.328-336
• Main Authors: Singh, Seema, Sadanandam, Anguraj, Varney, Michelle L., Nannuru, Kalyan C., Singh, Rakesh K.
• Format: Article
• Language: English
• Subjects: Actins - metabolism; Angiogenesis; Animals; Apoptosis; Apoptosis - genetics; Apoptosis - physiology; Blotting, Western; Cancer; Cell growth; Cell Line, Tumor; Cell migration; Cell Proliferation; Cell survival; Cell Survival - genetics; Cell Survival - physiology; chemokines; CXCR1; CXCR2; CXCR2 protein; Cytoskeleton; Endothelial cells; Female; Humans; Medical research; Melanoma; Melanoma - blood supply; Melanoma - genetics; Melanoma - pathology; Melanoma, Experimental - blood supply; Melanoma, Experimental - genetics; Melanoma, Experimental - pathology; Metastases; Mice; Mice, Nude; Mitogen-Activated Protein Kinase 1 - metabolism; Mitogen-Activated Protein Kinase 3 - metabolism; Neoplasm Invasiveness; Neoplasm Transplantation; Neovascularization, Pathologic - genetics; Neovascularization, Pathologic - pathology; Phosphorylation; Receptors, Interleukin-8A - genetics; Receptors, Interleukin-8A - metabolism; Receptors, Interleukin-8A - physiology; Receptors, Interleukin-8B - genetics; Receptors, Interleukin-8B - metabolism; Receptors, Interleukin-8B - physiology; RNA Interference; RNA, Small Interfering - genetics; siRNA; Skin cancer; Targeted cancer therapy; Transfection; Transplantation, Heterologous; Tumor Burden; tumor growth; Tumors; Xenografts
• ISSN: 0020-7136; 1097-0215
• DOI: 10.1002/ijc.24714

CXCR1 and CXCR2 are receptors for CXCL‐8 and are differentially expressed on melanoma and endothelial cells. In this study, we determined the functional role of these receptors in melanoma progression. We stably knock‐down the expression of CXCR1 and/or CXCR2 in A375‐SM (SM; high metastatic) human melanoma cells by short‐hairpin RNA transfection. Cell proliferation, migration, invasion, ERK phosphorlyation and cytoskeletal rearrangements were carried out in vitro. In vivo growth was evaluated using murine subcutaneous xenograft model. Our data demonstrate that knock‐down of CXCR1 and/or CXCR2 expression, inhibited melanoma cell proliferation, survival, migration and invasive potential in vitro. Moreover, we also observed inhibition of ERK phosphorylation and cytoskeltal rearrangement in SM‐shCXCR1, SM‐shCXCR2 and SM‐shCXCR1/2 cells. Furthermore, when SM‐shCXCR1 or SM‐shCXCR2 cells implanted in nude mice, tumor growth, proliferation and microvessel density was significantly inhibited as compared to SM‐control cells. In addition, we observed a significant increase in melanoma cell apoptosis in SM‐shCXCR1 and SM‐shCXCR2 tumors compared to SM‐control tumors. Together, these data demonstrate that CXCR1 and CXCR2 expression play a critical role in human melanoma tumor progression and, functional blockade of CXCR1 and CXCR2 could be potentially used for future therapeutic intervention in malignant melanoma.