Abstract
BackgroundGenome-wide RNA interference screening study revealed that loss of expression of insulin-like growth factor binding protein 7 (IGFBP7) is a critical step in development of a malignant melanoma (MM), and this secreted protein plays a central role in apoptosis of MM. In this study we constructed pcDNA3.1-IGFBP7 to obtain high expression of IGBPF7 and to inhibit the growth of MM in C57BL/6J mice.MethodspcDNA3.1-IGFBP7 was transfected into B16-F10 cell, the expression of IGFBP7 was detected by RT-PCR and western blot. The proliferations and apoptosis rates of transfected and control cells were measured by CCK8 and FCM, respectively. The tumorigenicity and tumor growth in both pcDNA3.1-IGFBP7 group and control groups were studied in C57BL/6J mice model. IGFBP7, caspase-3, and VEGF expressions in tumor tissue were measured by immunohistochemistry. Apoptosis of tumors were detected by TUNEL.ResultsWe demonstrated this plasmid inhibited proliferation of B16-F10 melanoma cells efficiently in vivo, exploiting the high expression of IGFBP7. More importantly, in-vivo transfection of pcDNA3.1-IGFBP7 inhibited MM growth in C57BL/6J mice. The inhibition of MM growth was proved owing to apoptosis and reduced expression of VEGF induced by pcDNA3.1-IGFBP7.ConclusionsThese results suggest a potential new clinical strategy for MM gene treatment.
Highlights
Genome-wide RNA interference screening study revealed that loss of expression of insulin-like growth factor binding protein 7 (IGFBP7) is a critical step in development of a malignant melanoma (MM), and this secreted protein plays a central role in apoptosis of MM
We demonstrated that the synthesis of vascular endothelial growth factor (VEGF) and growth of MM in xenograf models [3] were significantly inhibited by using small-interfering RNA, which makes us believe that the modulation of aberrant signaling pathways in MM cell will probably provide more effective and potential nontoxic therapy for MM
Identification of pcDNA3.1-IGFBP7 plasmid The sequence analysis of constructed pcDNA3.1IGFBP7 by a DNA sequencer showed the same sequence of eukaryotic IGFBP7 mRNA as designed
Summary
Genome-wide RNA interference screening study revealed that loss of expression of insulin-like growth factor binding protein 7 (IGFBP7) is a critical step in development of a malignant melanoma (MM), and this secreted protein plays a central role in apoptosis of MM. We demonstrated that the synthesis of vascular endothelial growth factor (VEGF) and growth of MM in xenograf models [3] were significantly inhibited by using small-interfering RNA (siRNA), which makes us believe that the modulation of aberrant signaling pathways in MM cell will probably provide more effective and potential nontoxic therapy for MM. This approach still has its shortcomings, in Recently, immunohistochemical analysis of human skin, nevi, and melanoma samples implicates loss of IGFBP7 expression as a critical step in melanoma carcinogenicity [9]. These exciting data suggested that IGFBP7 may be an efficacious anticancer agent, since experiments have provided evidences that IGFBPs have both IGF-dependent and IGF-independent antitumoral actions [13,14]
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