Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
5512223 | International Journal of Biological Macromolecules | 2017 | 17 Pages |
Abstract
Ovarian carcinoma is a tumor derived from ovary, which brings relatively higher mortality rate among the fatal gynecological cancers. Recently, lots of studies have concentrated on the anti-tumor effects of Arg-Gly-Asp (RGD) motif containing peptides due to their integrin binding properties. In order to meet the criterion of genetic engineering drugs, a recombinant RGD toxin protein (rLj-RGD3) without a His-tag was cloned from the buccal glands of Lampetra japonica in the present study. After endotoxin removal, the His-tag removed rLj-RGD3 was shown to inhibit the proliferation of HeyA8 cells. According to the confocal microscope, flow cytometry and western blot analysis, rLj-RGD3 could trigger HeyA8 cells apoptosis by changing mitochondrial membrane potential, arrangement of F-actin, protein level of BCL2, BAX, caspase 3, and cleaved caspase 3, concentration of cytoplasmic calcium, as well as phosphorylation level of ERK/JNK/p38. Furthermore, rLj-RGD3 was also able to suppress the adhesion, migration, and invasion processes of HeyA8 cells by disturbing the organization of F-actin and reducing the level of p-FAK. In addition, rLj-RGD3 could inhibit the adhesion of HeyA8 cells to extracellular matrix proteins by competitively binding to integrins, indicated that rLj-RGD3 might act as an anti-tumor drug to treat ovarian carcinoma patients in the future.
Keywords
ECMBCARh123ERKFAKRGDGAPDHDCFH-DAbFGFArg-Gly-AspPMSFORFrLj-RGD3TBSTPVDFFBSE. coliMMPPBSTris buffered saline TweenDMEML. japonicaJnkPTK2BSAc-Jun N-terminal kinasesDulbecco's modified Eagle MediumLampetra japonicaMAPKMitogen activated protein kinasesMTTROSbovine serum albuminEDTAEthylenediaminetetraacetic acidSprague Dawleymatrix-assisted laser desorption/ionization time-of-flight mass spectrometryEscherichia colisodium dodecyl sulfate-polyacrylamide gel electrophoresisSDS-PAGEbicinchoninic acidpolyvinylidene difluorideRhodamine123fetal bovine serumAnti-tumorbasic fibroblast growth factorPhosphate buffered salinePhenylmethanesulfonyl fluorideopen reading framelactate dehydrogenaseLDHExtracellular matrixMatrix metallo-proteinaseMALDI-TOF-MSprotein tyrosine kinase 2Propidium iodideoptical densityhigh performance liquid chromatographyHPLCcaspaseextracellular signal-regulated kinasefocal adhesion kinaseglyceraldehyde-3-phosphate dehydrogenaseReactive oxygen species
Related Topics
Life Sciences
Biochemistry, Genetics and Molecular Biology
Biochemistry
Authors
Qi Jiang, Qingwei Li, Jianmei Han, Meng Gou, Yuanyuan Zheng, Bowen Li, Rong Xiao, Jihong Wang,