1. In-vivo transfection of pcDNA3.1-IGFBP7 inhibits melanoma growth in mice through apoptosis induction and VEGF downexpression
- Author
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Yating Tu, Hongxiang Chen, Ping Jiang, Li Xu, Jia-Xi Lin, Wei-Bing She, and Rong-Yi Chen
- Subjects
Cancer Research ,DNA, Complementary ,Skin Neoplasms ,IGFBP7 ,Cell Survival ,medicine.medical_treatment ,Cell ,Melanoma, Experimental ,Apoptosis ,Transfection ,lcsh:RC254-282 ,Insulin-like growth factor-binding protein ,Mice ,Western blot ,In vivo ,medicine ,Animals ,biology ,medicine.diagnostic_test ,Vascular Endothelial Growth Factors ,Growth factor ,Research ,Genetic Therapy ,lcsh:Neoplasms. Tumors. Oncology. Including cancer and carcinogens ,Molecular biology ,Insulin-Like Growth Factor Binding Proteins ,medicine.anatomical_structure ,Oncology ,biology.protein ,Cancer research ,Plasmids - Abstract
Background 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. 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. Methods pcDNA3.1-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. Results We 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. Conclusions These results suggest a potential new clinical strategy for MM gene treatment.
- Published
- 2010