[Effect of PCDGF shRNA on growth, proliferation, apoptosis of human breast carcinoma cell line MCF-7 and expression of VEGF].
Tao Huang, Haiping Song, Jinghui Zhang
Abstract
Tao Huang, Haiping Song, Jinghui Zhang
Abstract
BACKGROUND & 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%. CONCLUSION: PCDGF regulates growth, proliferation, and apoptosis of MCF-7 cells and the expression of VEGF.
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BACKGROUND & 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%. CONCLUSION: PCDGF regulates growth, proliferation, and apoptosis of MCF-7 cells and the expression of VEGF.
Key concepts: Transfection, Apoptosis, MCF-7, Small hairpin RNA, Cell growth, Vascular endothelial growth factor, Cancer research, Biology