| Ovarian cancer is one of the popular malignant tumors of gynecology and its mortality is the highest of gynecological malignant tumors. Fibroblast growth factor (FGF) is a kind of growth factor with powerful mitogenic characteristics. FGF could facilitate cell growth and differentiation in many kinds of cells including fibroblast and is related to carcinogenesis. Acidic fibroblast growth factor ( aFGF) and basic fibroblast growth factor (bFGF) are two major factors of FGF family. Overexpression of FGF in renal cell cancer,urothelial cancer,prostate cancer,colonic cancer, pulmonary cancer and mammary cancer has been reported. Overexpression of bFGF and its receptor also has been reported in ovarian cancer. There is no report about aFGF and ovarian cancer up to day. We explored the expression of aFGF ?bFGF and their receptor FGFRj in 40 cases of o-varian epithelial cancer by RT - PCR and western blot.The transduction pathway of FGF in ovarian cancer is unknown up to day. We observed the effects of aFGF and bFGF on the proliferation of ovarian cancer cell line C AOV3 and the changes of activity of protein kinase C (PKC) and extracellular regulated protein kinase ( ERK) , and the effect of ERK kinase MEKt inhibitor PD98059 on the cell proliferation and activity of PKC and ERK.Materials and Methods1. Exprssion of aFGFmRNA,bFGFmRNA and FGFR1mRNA in ovarian epithelial cancer tissue were detected by RT - PCR; Expression of bFGF and FG-FR, in ovarian epithelial cancer tissue were detected by western blot.2. Exprssion of aFGF,bFGF and PD98059 on the proliferation of ovarian cancer cell line CAOV3 were detected by cytometry and MTT assay .3. Cell morphological changes induced by PD98059 were detected by inverted phase contrast microscope -, scanning electron microscope , transmission e-lectron microscope and Hoechst 33258 fluorescence.4. Apoptosis of CAOV3 cell induced by PD98059 was detected by DNA -ladder electrophoresis,Flow cytometry,Terminal deoxynucleotidyl tranferase mediated dUTP nick end labeling (TUNEL).5. The activity of PKC and ERK in CAOV3 cells induced by different concentration of aFGF,bFGF and PD98059 was detected by in corporation of [ r -32 P] - ATP into exogenous substrate.Results1. Expression of aFGF,bFGF and FGFR1 in ovarian epithelial cancer.There were overexpression of aFGF,bFGF and FGFR, in ovarian cancer and ovarian borderline tumors. Compared with normal ovary , ovarian tumor - like condition and ovarian benign tumors, the difference was significant ( P < 0. 05 ) . There were no difference of the expression of aFGF,bFGF and FGFR, in normal ovary, ovarian tumor - like condition and ovarian benign tumors. The expression of aFGF, bFGF and FGFRj in stage IE ~ IV was higher than that in stage I ~ II and ovarian borderline tumors ( P <0. 05) . There was no difference between stage I - It and ovarian borderline tumors. There were positive relationships in aFGF and bFGF.aFGF and FGFR, ,bFGF and FGFR, ( P <0. 05). There was no significant difference in differentiation and pathological types.2. The cell proliferative rate increased with aFGF and bFGF. When the concentration of bFGF was at 75ng/ml, the cell proliferative rate was at the peak. The proliferative rate decreased with PD98059 in a dose - dependent manner, especially in cells treated with aFGF and bFGF( P <0. 05).3. The morphological changes induced by PD98059.Under inverted phase contract microscope, the cell density obviously reduced , the cellular volume become smaller, cell got round and there was vacu-ole like structure in cell. Under scanning microscope, the cellular volume become smaller, the microvillus on the surface of cell disappeared. Under transmission microscope, the chromatin was condense and margined. Under Hoechst 33258 fluorescence, the apoptosis cells had densitive granules in nuclei.4. Degradation of the nuclei DNA of apoptosis cells was demonstrated by visualization of "DNA ladders" on gel electrophoresis. Moreover, DNA flow cy-tometry and TUNEL of DNA break... |
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