| Objective:Dextran sulfate(DS)is a macromolecular dextran derivative,which is considered to be a potential anticancer drug,but its role in the occurrence and development of ovarian cancer has not been studied.The purpose of this study was to observe the effect of DS on the malignant biological behavior of ovarian cancer cells(A2780 and SKOV3)and explore its possible mechanism.Methods:Ovarian cancer cell lines A2780 and SKOV3,were cultured in vitro and divided into control group and DS treatment group(DS concentration: 0.1%,0.3%,0.6%,1%).After treatment for 0 h,12 h,24 h and 48 h,respectively,the proliferation of ovarian cancer cell lines A2780 and SKOV3 were examined by CCK-8 cell growth experiment and colony-forming unit assay,and the maximal percentage inhibition was counted.The effects of control group and 1%DS treatment group on the transfer ability of ovarian carcinoma cells A2780 and SKOV3 were decided by wound healing assay and Transwell migration assay.The effects of control group and 1%DS treatment group on the invasiveness of ovarian cancer cells A2780 and SKOV3 were decided by Transwell invasion assay.The expression of PI3 K,p-PI3 K,AKT,p-AKT,mTOR,p-mTOR,E-cadherin,N-cadherin,Vimentin and Snail in ovarian cancer cells A2780 was examined by Western Blotting.Ovarian cancer cells A2780 and SKOV3,were again divided into control group,1%DS treatment group and 1%DS+SC79 group.The above functional experiments were repeated to verify the effects of DS on ovarian cancer proliferation,migration and invasion and the changes of key protein expression.Results:1.CCK-8 assay explained that with the raise of the action time and drug concentration of DS on ovarian cancer cells(A2780 and SKOV3),the inhibitory of DS on the multiplication capacity of ovarian cancer cells became stronger,it was time-and concentration-dependent.When the drug concentration was 0.1%,the inhibition rate were6.13%、5.79% at 12 h and 21.98%、20.31% at 48 h.When the drug concentration was 1.0%,the inhibition rate were 40.2% 、 40.72% at 12 h and 57.51% 、 58.79% at 48 h.The difference was statistically significant(P < 0.001).2.Colony formation assay showed that the number of clones in DS treated group(A2780 cells: 593.7 ±8.99;SKOV3 cells: 167.3 ±4.81)was significantly lower than that in the control group(A2780 cells: 530.3 ±14.1;SKOV3 cells: 163 ±6.08)(P < 0.001).3.Wound healing assay showed that after 48 hours of treatment in the control group and 1%DS treatment group,the healing rates of A2780 cells and SKOV3 cells were74.02%,46.84% and 63.5%,45.82% respectively(P < 0.001).4.The results of Transwell assay showed that after 48 hours of treatment of A2780 cells and SKOV3 cells in control group and 1%DS treatment group,the number of migrating cells was(366.7 ±4.485),(183.3 ±5.783)and(152.7 ±6.642),respectively(P <0.001).After 48 hours of treatment with A2780 cells and SKOV3 cells in control group and 1%DS treatment group,the number of invasive cells were(353.3 ±12.91,161 ±9.292)and(350 ±11.85,169.3 ±16.95),respectively(P < 0.001).5.The results of Western Blotting showed that there was no obvious change in the expression of PI3 K,P-PI3 K,AKT and mTOR proteins in the 1%DS treatment group compared with the control group.The expression levels of p-AKT,p-mTOR,N-cadherin,E-cadherin,Vimentin and Snail in the 1%DS treated group were 0.1281 ±0.0172,0.2386±0.0041,0.1575 ±0.0201,0.4123 ±0.0268,0.0797 ±0.0012 and 0.1471 ±0.0019,respectively,compared with those in the control group(0.2848 ±0.0169,0.5032±0.0098,0.2247 ±0.0241,0.2668 ±0.045,0.1091 ±0.02,0.2504 ±0.0203),the difference was statistically significant(P < 0.05).6.Functional recovery test: the addition of AKT agonist(SC79)reversed the inhibitory effect of DS on the proliferation,migration and invasion of A2780 and SKOV3 cells.Western Blotting showed that the inhibitory effect of DS on AKT phosphorylation was reversed by adding SC79.Conclusion1.The proliferation,migration and invasion of DS ovarian cancer cells were inhibited.2.DS can inhibit epithelial-mesenchymal transformation(EMT))of ovarian cancer cells.3.DS inhibits the malignant behavior of ovarian cancer cells by inhibiting PI3K/AKT /mTOR signal pathway. |
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