The Effects Of Recombinant Human IL-17D On Proliferation,migration And Expression Of VEGF-C In Human Ovarian Cancer Cell Line SKOV3

Objectives: This thesis investigates the effects of recombinant human IL-17 D on ovarian cancer proliferation and lymphatic metastasis,by studying the effects of recombinant human IL-17 D on the proliferation,migration and expression of VEGF-C in human ovarian cancer SKOV3 cells.Methods:1 The human ovarian papillary serous adenocarcinoma cell lines of SKOV3 were cultured in vitro,and observed cells morphological by inverted microscope.2 Collect SKOV3 cells in logarithmic growth phase.Use the MTS to test the proliferation of SKOV3 cells cultured with different concentration of r-hIL-17D(0 ng/ml,10 ng/ml,50 ng/ml,100 ng/ml)at different time points(0,12,24,48,72,96,120h),and draw the growth curve of cells.3 Use Transwells to assay the migration ability of SKOV3 cells influenced by different concentrations of r-hIL-17D(0 ng/ml,10 ng/ml,50 ng/ml,100 ng/ml).4 Isolate the RNA of the SKOV3 cells cultured with different concentration of r-h IL-17D(0 ng/ml,10 ng/ml,50 ng/ml,100 ng/ml)for 48 h.RT-q PCR was used to detect the expression of VEGF-C in SKOV3 cells.5 SKOV3 cells were cultured with different concentration of r-hIL-17D(0 ng/ml,10 ng/ml,50 ng/ml,100 ng/ml)for 48 h,and the content of VEGF-C in cell supernatant was detected by ELISA method.Result:1 The SKOV3 cells were successfully cultured and passaged,and the logarithmic growth phase cells were obtained.2 MTS method showed the proliferation rates of SKOV3 cells in 10 ng/ml,50 ng/ml,100 ng/ml group were faster than 0 ng/ml group,and the difference was statistically significant(P <0.05).The proliferation rate of SKOV3 cells in 50 ng/ml,100 ng/ml group was faster than 10ng/ml group,the difference was statistically significant(P <0.05).There was no significant difference between 50 ng / ml group and 100 ng / ml group on the aspect of proliferation rate(P>0.05).3 The Transwell assay showed that the number of SKOV3 cells migration,which was cultured with different concentrations of r-hIL-17D(0 ng / ml,10 ng / ml,50 ng / ml,100 ng / ml),were 118.80 ± 13.66,144.20±12.670,180.80±20.54,176.60±18.15.The difference was statistically significant(P <0.05).The migration ability of SKOV3 cells in 10 ng / ml,50 ng / ml and 100 ng / ml groups was stronger than that in 0 ng / ml group(P <0.05).The migration ability of SKOV3 cells in 50 ng / ml and 100 ng / ml groups was stronger than that in 10 ng / ml group(P <0.05).There was no significant difference between 50 ng / ml group and 100 ng / ml group on migration ability(P>0.05).4 The quantization of relative expression of VEGF-C mRNA in SKOV3 cells cultured with different concentrations of r-hIL-17D(0 ng / ml,10 ng / ml,50 ng / ml,100 ng / ml)were 1.00 ± 0.18,1.54 ± 0.22,2.26 ± 0.18,2.11 ± 0.26,and the difference was statistically significant(P <0.05).The expression of VEGF-C mRNA of SKOV3 cells in 10 ng / ml,50 ng / ml and 100 ng / ml groups was higher than that in 0 ng / ml group(P <0.05).The expression of VEGF-C mRNA of SKOV3 cells in 50 ng / ml and 100 ng / ml groups was also higher than that in 10 ng / ml group(P <0.05).There was no significant difference between 50 ng / ml group and 100 ng / ml group(P>0.05).5 The results of ELISA showed:the levels of VEGF-C in the supernatants of SKOV3 cells cultured with different concentrations of r-hIL-17D(0 ng / ml,10 ng / ml,50 ng / ml,100 ng / ml)respectively were 4095.09 ± 105.25 ng / ml,4439.84 ± 133.87 ng / ml,4755.66 ± 64 ng / ml,4646.59 ± 111.92 ng / ml,and the difference was statistically significant(P <0.05).The levels of VEGF-C in the supernatants of SKOV3 cells in 10 ng / ml,50 ng / ml and 100 ng / ml groups were higher than that in 0 ng / ml group(P <0.05).The levels of VEGF-C in the supernatants of SKOV3 cells in 50 ng / ml and 100 ng / ml groups were also higher than that in 10 ng / ml group(P <0.05).There was no significant difference between 50 ng / ml group and 100 ng / ml group(P>0.05).Conclusion:R-hIL-17 D can promote the proliferation of SKOV3 cells,enhance the migration ability of SKOV3 cells and increase the expression of VEGF-C of SKOV3 cells.IL-17 D may promote metastasis of ovarian cancer by those ways.