[Effect of PCDGF shRNA on growth, proliferation, apoptosis of human breast carcinoma cell line MCF-7 and expression of VEGF].
OBJECTIVE
PC cell-derived growth factor (PCDGF) is a crucial factor for regulating the growth and apoptosis of breast cancer cells; however the mechanism is unclear. This study was to construct a short hairpin RNA (shRNA) targeting PCDGF, and analyze its effects on growth, proliferation, and apoptosis of MCF-7 cells and the expression of vascular endothelial growth factor (VEGF).


METHODS
The expression of PCDGF in breast cancer cell lines MCF-7, T47D, MDA-MB-435, and MDA-MB-231 was detected by reverse transcription-polymerase chain reaction (RT-PCR). Two shRNAs targeting PCDGF were combined to pGenesil-1 vector, and transfected into MCF-7 cells. Cell growth and proliferation were analyzed by MTT assay; VEGF expression was detected by ELISA.


RESULTS
PCDGF was diversely expressed in these 4 cell lines. After transfection of PCDGF shRNA, the inhibitory rate of PCDGF expression in MCF-7 cells was 59.8%; the apoptosis rates were 30.41% at early phase and 35.38% at late phase in PCDGF shRNA-transfected MCF-7 cells, and were 3.69% at early phase and 15.39% at late phase in control cells. The inhibitory rate of VEGF expression in PCDGF shRNA-transfected MCF-7 cells was 47.2%.


CONCLUSIONS
PCDGF regulates growth, proliferation, and apoptosis of MCF-7 cells and the expression of VEGF.
Surgical Center of Thyroid and Breast, Union Hospital, Huazhong University of Science and Technology, Wuhan, Hubei 430022, P. R. China. whshp@163.com
Tao
T
Huang
Hai-Ping
HP
Song
Jing-Hui
JH
Zhang
D017209:Apoptosis; D001943:Breast Neoplasms / Q000378:metabolism / Q000473:pathology*; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D005260:Female; D005822:Genetic Vectors; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000096:biosynthesis* / Q000235:genetics; D010957:Plasmids; D000077153:Progranulins; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics*; D014162:Transfection; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism*
C492314:GRN protein, human; D036341:Intercellular Signaling Peptides and Proteins; D000077153:Progranulins; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D042461:Vascular Endothelial Growth Factor A