| Backgroud and ObjectiveOvarian cancer is one of the most common malignant tumors in female genitalia.Due to the early nonspecific clinical symptoms,the vast majority of patients have entered the advanced stage when they are diagnosed,with a very low five-year survival rate.Epithelial ovarian cancer accounts for about 90%of primary ovarian cancer,its occurrence and the mechanisms of development is complex.Vascular endothelial growth factor(VEGF),platelet derived growth factor(PDGF)can participate in many tumor processes by promoting angiogenesis in tumor tissues.Neovascularization provides the necessary nutritional support for the growth of tumor cells.Tumor growth,migration and metastasis and clinical grade can be associated with angiogenesis.However,the expression of pro-angiogenic factors in ovarian cancer,especially in patients with epithelial ovarian cancer,how they participate in the development of epithelial ovarian cancer remains to be further explored.In the early stage,we found that peripheral blood PDGF levels increased in patients with epithelial ovarian cancer,and on this basis,we further detected serum PDGF levels in patients with epithelial ovarian cancer in different clinical stages by enzyme-linked immunosorbent assay(ELISA).The expression of PDGF mRNA in epithelial ovarian cancer tissues and adjacent normal tissues was detected by real-time quantitative PCR(qPCR).The expression of PDGF protein in epithelial ovarian cancer tissues and adjacent normal tissues was detected by immunohistochemistry.In order to verify the experimental results,we used ovarian epithelial cancer cell line OVCAR-3 as a model to supplement PDGF with different concentrations in vitro.The effects of PDGF on the migration and proliferation of OVCAR-3 were detected by Transwell assay and MTT assay.Material and Methods1.Serum PDGF levels were detected by enzyme-linked immunosorbent assay(ELISA)in 20 cases of control subjects and 12 cases of stage I,16 cases of stage II,18 cases of stage III,21 cases of stage IV ovarian epithelial cancer,and the correlation of the serum PDGF levels and ovarian epithelial cancer clinical stage were analyzed.2.Real-time quantitative PCR(qPCR)and immunohistochemistry were used to detect the expression of PDGF mRNA and protein,respectively,in 25 cases of ovarian cancer tissues and adjacent tissues.3.The ovarian epithelial cancer cell line OVCAR-3 was used as a model to supplement different concentrations of PDGF in vitro.The effects of PDGF on the migration and proliferation of OVCAR-3 were detected by Transwell assay and MTT assay.Results1.The serum PDGF levels in the healthy control group were(118.95±25.31)pg/ml,the PDGF concentrations in the ovarian epithelial cancer of stage I-IV were(168.50±23.11)pg/ml,(193.06±22.34)pg/ml,(202.61±23.62)pg/ml,(249.57±32.41)pg/ml,respectively.The PDGF concentrations of healthy control was lower than that of the patients with epithelial ovarian cancer in each stage,and there was a significant difference(p<0.05).With the clinical stage of ovarian epithelial cancer increased,serum PDGF content showed an upward trend.2 Compared with the healthy control group,PDGF levels in peripheral blood of patients with epithelial ovarian cancer not only increased with the development of clinical stage,but also corrected with the clinical stage(r=0.859,p<0.0001).3 The relative expression of PDGF mRNA in para-cancer tissue was(1349.32±203.64)folds,while the relative expression level of PDGF mRNA in cancer tissue was(1664.20±187.41)folds.The average age of patients with epithelial ovarian cancer was(52.84±11.73).Paired t-test analysis showed that the relative expression of PDGF mRNA in epithelial ovarian cancer patients was significantly higher than that in paracancerous tissues(p<0.0001).4.The characteristic of ovarian epithelial cancer tissue observed via the microscopic showed that neovascular were common,vascular endothelial cells were monolayer,thin wall,and blood cells filled with the official cavity.Cancer cells were clustered to form a germinal center.The cancer cells with highly heterogeneous nuclei,abundant cytoplasm,large nuclei and loose chromatin were common,suggesting that the cancer were highly proliferative state.5.The average density of PDGF positive signal in adjacent tissues was(0.27±0.12)C,and the average density of PDGF positive signals in cancer tissues was(1.63±0.43)C.Paired t test showed that PDGF protein expression in cancer tissue of patients with epithelial ovarian cancer was significantly higher than that in the para-cancerous tissue(p<0.001).6.The number of OVCAR-3 cell that crossing the bio-membrane were(0.88±0.13)/mm~2 in the group without PDGF protein,while the number in low PDGF protein group and high PDGF protein group was(1.40±0.18)/mm~2 and(2.03±0.33)/mm~2,respectively.Analysis of variance showed that,compared with the blank group,low and high concentration of PDGF could effectively induce cell migrate through the bio-membrane(both p<0.05),which would dependent on the concentration of PDGF.7.The OD490 absorbance value was(0.19±0.07)in the group without PDGF protein,while the value in low PDGF protein group and high PDGF protein group was(1.10±0.17)and(2.37±0.26),respectively.The analysis of variance showed that the OD490 absorbance values were both higher in low and high concentration of PDGF group(both p<0.0001).Conclusion1.Serum PDGF levels in peripheral blood of patients with epithelial ovarian cancer were significantly increased and positively correlated with the stage of the disease.The expression of PDGF mRNA and protein in cancer tissues was higher than that in para-cancerous tissues.2.In vitro,PDGF can promote the proliferation and migration of ovarian cancer cell line OVCAR-3 in a concentration-dependent manner. |
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