Proteomics reveals protein profile changes in cyclooxygenase-2 inhibitor-treated endometrial cancer cells.
OBJECTIVE
To examine effects of an inhibitor of cyclooxygenase (COX)-2, NS-398, on the proliferation, apoptosis and invasion characteristics of endometrial cancer cell RL95-2.


METHODS
(1) Western blotting was carried out to determine COX-2 protein expression in RL95-2 cells and normal endometrium specimens. (2) The effect of NS-398 treatment on the cell proliferation, apoptosis, and invasion was assessed by methyl thiazolyl tetrazolium assay, flow cytometry, and matrigel invasion assay, respectively. (3) Finally, the proteomic analysis was used to find out proteins that are differentially expressed because of NS-398 treatment.


RESULTS
(1) COX-2 protein in RL95-2 cell line was significantly higher than that in normal endometrium. (2) NS-398 had significant growth inhibition effects on RL95-2 cells in a dose- and time-dependent manner. (3) NS-398 increased the proportion of cells in G1 and decreased the proportion of cells in the G2 phase in RL95-2 cells. (4) NS-398 could restrain endometrial cancer cells invasion. (5) The proteomic analysis revealed several proteins that are differentially expressed because of NS-398 treatment; the down-regulated proteins identified are hnRNP K, alpha enolase, Hsp70, tropomyosin, and protein disulfide isomerase, the up-regulated protein is phosphatidylethanolamine binding protein.


CONCLUSIONS
The expression of COX-2 plays an important role in tumorigenesis of endometrial cancer. NS-398 can inhibit the ability of RL95-2 cell proliferation, viability, and invasion. In this study, the well-resolved reproducible 2-DE maps of NS-398 treated and control RL95-2 cells were established, and the significantly different expressed proteins are preliminary identified.
Yi
Zhang
Z
Department of Obstetrics and Gynecology, Xiangya Hospital, Central South University, Changsha, Hunan, People's Republic of China.
Jingting
Cai
C
Yu
Zhang
Z
D017209:Apoptosis / Q000187:drug effects; D014408:Biomarkers, Tumor / Q000378:metabolism; D015153:Blotting, Western; D002448:Cell Adhesion / Q000187:drug effects; D002453:Cell Cycle / Q000187:drug effects; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D051546:Cyclooxygenase 2 / Q000737:chemistry / Q000378:metabolism*; D052246:Cyclooxygenase 2 Inhibitors / Q000494:pharmacology*; D015180:Electrophoresis, Gel, Two-Dimensional; D016889:Endometrial Neoplasms / Q000188:drug therapy / Q000378:metabolism* / Q000473:pathology; D004717:Endometrium / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D005260:Female; D015971:Gene Expression Regulation, Enzymologic; D006801:Humans; D009578:Nitrobenzenes / Q000494:pharmacology*; D040901:Proteomics; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013449:Sulfonamides / Q000494:pharmacology*; D014407:Tumor Cells, Cultured
D014408:Biomarkers, Tumor; D052246:Cyclooxygenase 2 Inhibitors; D009578:Nitrobenzenes; D013449:Sulfonamides; C080955:N-(2-cyclohexyloxy-4-nitrophenyl)methanesulfonamide; D051546:Cyclooxygenase 2; C496540:PTGS2 protein, human