In-vivo transfection of pcDNA3.1-IGFBP7 inhibits melanoma growth in mice through apoptosis induction and VEGF downexpression.
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.
Department of Dermatology, Affiliated Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022 PR China.
Rong-Yi
RY
Chen
Hong-Xiang
HX
Chen
Jia-Xi
JX
Lin
Wei-Bing
WB
She
Ping
P
Jiang
Li
L
Xu
Ya-Ting
YT
Tu
D000818:Animals; D017209:Apoptosis; D002470:Cell Survival; D018076:DNA, Complementary; D015316:Genetic Therapy; D018969:Insulin-Like Growth Factor Binding Proteins / Q000235:genetics*; D008546:Melanoma, Experimental / Q000628:therapy*; D051379:Mice; D010957:Plasmids; D012878:Skin Neoplasms / Q000628:therapy*; D014162:Transfection; D042442:Vascular Endothelial Growth Factors / Q000378:metabolism*
D018076:DNA, Complementary; D018969:Insulin-Like Growth Factor Binding Proteins; D042442:Vascular Endothelial Growth Factors; C102529:insulin-like growth factor binding protein-related protein 1