ABSTRACT
Objective:To investigate the molecular mechanism of PDGFC regulated by HuR in breast cancer cells.Methods:Through the software analysises,we first predicted the HuR-binding sites on the PDGFC 3'-UTR in breast cancer cells;An RNA-immunoprecipitation tested the interaction of HuR with the PDGFC mRN after adding PDGFC stimulation;Luciferase experiments tested the HuR-binding sites of PDGFC regulated by HuR through structuring five truncated of PDGFC 3'UTR.Results:We found five HuR-binding sites in the 3'UTR of PDGFC by software prediction;The RNA-immunoprecipitation showed the co-immunoprecipitation of HuR and PDGFC mRN after adding PDGFC stimulation confirming the direct association of HuR with the PDGFC;Luciferase experiments ofPDGFC mRNA 3'UTR showed that PDGFC regulated by the second and fourth HuR-binding sites.Conclusions:This study reveals the molecular mechanism of PDGFC regulated by HuR through binding to PDGFC mRNA 3'UTR in breast cancer cells,and provided rationale for the development of strategies in the clinical diagnosis and treatment for breast cancer.
ABSTRACT
Objective:Abnormal angiogenesis is an important hallmark of HCC. Ectopic miR-375 overexpression led to repression of proliferation, migration, invasion, and colony formation, and it induced apoptosis in hepatoma cells as well. In this study, we explored the effect of miR-375 on HCC angiogenesis. Methods:We evaluated the antiangiogenic effects of miR-375 using human umbilical vein endothelial cells, tube formation assays, rat aortic ring sprouting assays, and chicken chorioallantoic membrane assays. Bioinformatics software was used to predict the downstream target gene of miR-375. MiR-375 regulation to target genes was explored by overexpres-sion and knockdown of miR-375 in hepatoma cells. Luciferase assay was performed to confirm its molecular mechanism. Rescue assay of target gene was further used to prove that miR-375 inhibited HCC angiogenesis by directly regulating its target gene. Results:MiR-375 inhibited HCC angiogenesis. Platelet-derived growth factor-C (PDGFC) was a potential target gene of miR-375. MiR-375 inhibited PDGFC expression in hepatoma cells by targeting its 3′-UTR. MiR-375 exerted its antiangiogenic effect partially by PDGFC inhibition. Conclusion:MiR-375 repressed tumor angiogenesis by targeting PDGFC in HCC.
ABSTRACT
BACKGROUND: Four platelet derived growth factor (PDGF) family members have been identified; the classical PDGFs, PDGF-A and PDGF-B, and the novel PDGFs, PDGF-C and PDGF-D, which were only recently discovered. METHODS: The present study was designed to determine the changes of the platelet derived growth factor (PDGF) subtypes (C & D) and their receptors (PDGFR)-alpha & beta expression in kidneys during pre- and postnatal development. RESULTS: All the protein levels of PDGFR-alpha and -beta and the mRNA levels of PDGF-C and D were high in kidneys during the prenatal period and decreased differently during the postnatal period. PDGFR-alpha was expressed in the interstitial space at embryo day 18. PDGFR-beta protein were expressed in metanephric blastema at embryo day 18. PDGF-C mRNA was expressed in metanephric blastema, developing glomerulus at embryo 18 day and in collecting duct at postnatal day 7. PDGF-D mRNA was expressed in the parietal and vesceral epithelial cells during pre and postnatal period. CONCLUSION: These results indicate that the PDGF subtypes (C & D) and their receptors (PDGFR-alpha & -beta) are differently expressed in the kidney during the prenatal and postnatal period.