ABSTRACT
Objective@#To explore the effect of the AXL expression on the chemosensitivity of prostate cancer PC-3 and DU145 cells to docetaxel and possible mechanisms.@*METHODS@#Using Western blot, we examined the expressions of the AXL protein, p-AXL and Gas6 in the docetaxel-resistant PC-3 (PC-3-DR) and DU145 (DU145-DR) cells stimulated with gradually increased concentrations of docetaxel. We transfected the PC-3 and DU145 cells with negative NC ShRNA and AXL-ShRNA, respectively, which were confirmed to be effective, detected the proliferation, apoptosis and cycle distribution of the cells by CCK8, MTT and flow cytometry after treated with the AXL-inhibitor MP470 and/or docetaxel, and determined the expression of the ABCB1 protein in the PC-3-DR and DU145-DR cells after intervention with the AXL-inhibitor R428 and/or docetaxel.@*RESULTS@#The expression of the AXL protein in the PC-3 and DU145 cells was significantly increased after docetaxel treatment (P <0.05). The expressions AXL and p-AXL were remarkably higher (P <0.05) while that of Gas6 markedly lower (P <0.05) in the PC-3 and DU145 than in the PC-3-DR and DU145-DR cells. The inhibitory effect of docetaxel on the proliferation and its enhancing effect on the apoptosis of the PC-3 and DU145 cells were significantly decreased at 48 hours after AXL transfection (P <0.05). MP470 obviously suppressed the growth and promoted the apoptosis of the PC-3-DR and DU145-DR cells, with a higher percentage of the cells in the G2/M phase when combined with docetaxel than used alone (P <0.05). R428 markedly reduced the expression of ABCB1 in the PC-3-DR and DU145-DR cells, even more significantly in combination with docetaxel than used alone (P <0.05).@*CONCLUSIONS@#The elevated expression of AXL enhances the docetaxel-resistance of PC-3 and DU145 prostate cancer cells and AXL intervention improves their chemosensitivity to docetaxel, which may be associated with the increased cell apoptosis in the G2/M phase and decreased expression of ABCB1.
Subject(s)
Humans , Male , ATP Binding Cassette Transporter, Subfamily B, Member 1 , Metabolism , Antineoplastic Agents , Pharmacology , Apoptosis , Cell Count , Cell Cycle , Cell Line, Tumor , Cell Proliferation , Docetaxel , Drug Resistance, Neoplasm , Intercellular Signaling Peptides and Proteins , Metabolism , Prostatic Neoplasms , Drug Therapy , Metabolism , Pathology , Proto-Oncogene Proteins , Genetics , Metabolism , Pyrimidines , Pharmacology , RNA, Small Interfering , Receptor Protein-Tyrosine Kinases , Genetics , Metabolism , Taxoids , PharmacologyABSTRACT
<p><b>OBJECTIVE</b>To investigate the effect of PI-3K and p38MAPK signal pathways on the cyclooxygenase-2 (COX-2) expression induced by the epidermal growth factor (EGF) in PC-3 cells.</p><p><b>METHODS</b>PC-3 cell proliferation was detected by methylthiazolyl tetrazolium (MTT) assay after stimulated by EGF (0 microg/L), EGF (10 microg/L), EGF (10 microg/L) + LY294002 (20 micromol/L) and EGF (10 microg/L) + SC203580 (20 micromol/L), and so was the COX-2 expression in the PC-3 cells by RT-PCR and Western blot assay after stimulated the same way for 24 hours. ELISA was used to determine the changes of PGE2 in the culture medium.</p><p><b>RESULTS</b>LY294002 and SC203580 signficantly inhibited PC-3 cell proliferation (P < 0.05), COX-2 expression and PGE2 production after EGF stimulation (P < 0.05).</p><p><b>CONCLUSION</b>EGF can stimulate PC-3 cells into proliferation and induce COX-2 mRNA and the upregulation of its protein expression, while LY294002 and SC203580 can inhibit EGF from the above effects on PC-3 cells.</p>
Subject(s)
Humans , Male , Blotting, Western , Cell Line, Tumor , Cell Proliferation , Cyclooxygenase 2 , Genetics , Metabolism , Dinoprostone , Metabolism , Enzyme-Linked Immunosorbent Assay , Epidermal Growth Factor , Pharmacology , Gene Expression , Phosphatidylinositol 3-Kinases , Metabolism , Reverse Transcriptase Polymerase Chain Reaction , Signal Transduction , p38 Mitogen-Activated Protein Kinases , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To investigate the changes of antisperm antibodies (AsAb), sexual hormones, and inhibin B (INH B) in patients before and after testicular torsion, as well as the effects of these factors on testicular function and reproduction.</p><p><b>METHODS</b>Ten patients with single acute testicular torsion (left side 9 and right side 1), aged 16-45 years (19.6 on average), disease course of 3-6 days (averaging 4.7 days), underwent surgical removal of the damaged testis. Before and after the operation, serum AsAb (IgG, IgM, IgA) and INH B were measured by ELISA, and serum follicle-stimulating hormone (FSH), luteotropic hormone (LH), and testosterone (T) determined by chemoluminescence autoanalyzer.</p><p><b>RESULTS</b>After the operation, the AsAb levels rose significantly and remained high for at least 26 weeks. The level of INH B was the lowest in the 3rd week and restored to normal in the 12th week, with significant difference between preoperation and the 3rd or the 6th week after the operation. The levels of LH and INH B in the 26th week were elevated significantly compared with the 6th.</p><p><b>CONCLUSION</b>Testicular injury induced the elevation of AsAb, which would last a very long time. The change of INH B was closely related with the injury of the testis, which reflected the degree of testicular injury and functional restoration of the patients after the operation. Our study showed that AsAb and INH B can be used as useful tools for monitoring testicular function and reproduction.</p>