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To assess roles of vascular endothelial growth factor (VEGF) and platelet-derived growth factor (PDGF) in the mechanisms of lymphangiogenesis in epithelial ovarian carcinoma.(1) Expression of Prox1, a newly described lymphatic endothelial cell nucleus marker, VEGF-A, VEGF-C, VEGF-D and PDGF-A, PDGF-B, PDGF-C, PDGF-D were detected by RT-PCR in SKOV3 cell line and in 90 ovarian tissue samples, included 15 benign tumors, 10 borderline tumors, 45 malignant tumors and 20 normal ovarian samples. (2) Expression levels of Prox1, VEGF-A, -C, -D and PDGF-A, -B, -C, -D were detected in 90 ovarian tissue sample mentioned above by real-time quantitative PCR (RTQ-PCR).(1) Prox1 was expressed in ovarian samples mentioned above, while not detected in SKOV3 cell. VEGF-A, -C, -D and PDGF-A, -B, -C, -D were found in SKOV3 cell and various ovarian tissues. (2) Expression levels of Prox1 (2.2 +/- 1.3, P0.01), VEGF-A (3.5 +/- 1.5, P0.01), VEGF-C (19 +/- 14, P0.01), VEGF-D (3.0 +/- 1.8, P0.01) and PDGF-A (3.3 +/- 3.3, P0.05), PDGF-C (6.9 +/- 4.6, P0.01) in malignant group were found to be significantly higher than those in borderline group and benign group. (3) The expression levels of Prox1, VEGF-A and PDGF-A were significantly greater in samples from the patients with lymph node metastasis (Prox1: 3.0 +/- 1.4, VEGF-A: 4.1 +/- 1.7, PDGF-A: 4.9 +/- 4.1), peritoneum metastasis (Prox1: 2.8 +/- 0.9, VEGF-A: 4.0 +/- 1.8, PDGF-A: 4.5 +/- 4.0) and in stage III - IV (Prox1: 2.6 +/- 1.3, VEGF-A: 4.0 +/- 1.4, PDGF-A: 4.1 +/- 3.7) than those without lymph node metastasis, without peritoneum metastasis and in stage I - II. There was a significant increased in the degree of VEGF-C and VEGF-D expression in positive lymph node metastasis group (VEGF-C: 24 +/- 13, VEGF-D: 3.9 +/- 2.0) compared with negative group (P0.05). (4) There were significant positive correlations between the expression levels of Prox1 and VEGF-D (r = 0.62, P0.01), PDGF-C (r = 0.91, P0.01) or PDGF-D (r = 0.61, P0.01).VEGF-A, VEGF-C and PDGF-A may promote lymphatic metastasis in epithelial ovarian carcinoma through else mechanisms other than lymphangiogenesis. VEGF-D may facilitate lymphangiogenesis and lymph node metastasis in epithelial ovarian cancer. There is no significant correlation between the expression of PDGF-B and lymphangiogenesis and lymph node metastasis. PCGF-C and PDGF-D may motivate lymphangiogenesis, but could not participate in lymph node metastasis in ovarian carcinoma.

To evaluate the influence of bcl-2 antisense oligodeoxynucleotides (AODN) on reversing cisplatin (DDP) resistance in ovarian carcinoma cell lines.In the experiment groups, cisplatin resistant ovarian carcinoma cell line A2780/DDP cells were treated with cisplatin after oligodeoxynucleotides were transfected by lipofectamine. Cell proliferation was determined by counting the number of the cells continuously for six days. The expression level of bcl-2 mRNA was detected with reverse transcription polymerase chain reaction (RT-PCR) and protein was tested by immunofluorescence technique. The apoptosis of ovarian cancer cell lines was measured by DNA ladder assay. The inhibitory rate of cells was detected by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT). The cells treated with culture media were used as the negative control, and the untreated cells were used as the blank.The proliferation of AODN transfected cells was inhibited significantly compared with that in the control groups, and the logarithmic growth delayed for 24h. The expression of bcl-2 mRNA and bcl-2 protein in AODN transfected cells was 0.76 +/- 0.01 and 171.3 +/- 3.1 respectively. Compared with the expression of bcl-2 mRNA 1.56 +/- 0.03 and bcl-2 protein 105.3 +/- 1.7 in the blank, the difference was significant (P0.01), but compared with the same concentration of sense oligodeoxynucleotides (SODN) or random-oligodeoxynucleotides (RODN) transfected cells, the difference was not significant (P0.05). The AODN transfected cells were sensitive to DDP again, and the inhibitory rate was (97.49 +/- 1.00)%, which was higher than that of the negative control (P0.01). In the cells treated with 16 microg/ml AODN for 72 h followed by DDP, DNA ladder was obviously observed with agarose gel electrophoresis, but in the RODN and SODN transfected cells, DNA ladder could not be seen.Through inhibiting the expression of bcl-2 mRNA and protein and increasing the apoptosis, AODN can reverse the resistance to cisplatin in ovarian cancer cell line A2780/DDP.

To construct an adenoviral vector in which the fusion gene cytosine and uracil phosphoribosyl (UPP) transferase was directed by glutathione S-transferase P1 (GSTP1) promoter, and to investigate specific killing effect of the suicide gene system on cisplatin-resistant ovarian cancer cells.Recombinant adenovirus was generated through homologous recombination in bacteria. A2780 and A2780/DDP cells were infected with Ad and then received flucytosine (5-FC) administration. The relative survival of these cells was tested. And a bystander effect was observed by mixing gene-transferred and gene-untransferred A2780/DDP cells with 5-FC.In vitro, when MOI was 100 and 5-FC was 250 micro g/ml, relative survival rate of A2780/DDP cells was only (3.6 +/- 1.0)%; that of A2780 cells was (76.5 +/- 2.8)%. Significant bystander effect was caused by CD-UPP gene and 20% gene-transferred A2780/DDP cells induced 80.3% of total cells to death.Recombinant adenovirus carrying CD-UPP gene driven by GSTP1 promoter has a specific killing effect on cisplatin-resistant ovarian cancer cells.