Effects Of Deuterium Depleted Water On Proliferation,Migration And Invasion Of Ovarian Cancer SKOV3 Cells

Objective: We used different concentration of deuterium depleted water(DDW)to act on human ovarian cancer SKOV3 cells in order to observe the effect of the proliferation,migration and invasion,and to explore its possible mechanism.To find out that DDW can become a scientific basis for the treatment of ovarian cancer.Methods:1.SKOV3 cells were cultured in vitro and divided into 4 groups.The experimental group used DDW medium with deuterium content of 125 ppm,75ppm,25 ppm,the control group was treated with three distilled water mediums with deuterium content of 150 ppm.2.Colony formation assay and CCK8 assay were used to determine the effect of DDW on the proliferation of SKOV3 cells at 24,48,72 and 96 hours.3.Transwell chamber migration test and scratch repair test was used to observe the effect of DDW on the migration ability of SKOV3 cells.4.Transwell chamber invasion test was used to observe the effect of DDW on the invasive ability of SKOV3 cells.5.Western blot was used to measure the expression of and matrix metallo protein 9(MMP-9)and vimentin(VIM)in SKOV3 cell.Results:1.The results of colony formation experiment showed that the number of cells in the control group was significantly more than that in the experimental group,and the number of cloned cells in the experimental group decreased with the decrease of deuterium concentration in the culture medium.However,after the analysis of statistical software,there was no statistical significance between the experimental group and the control group(P > 0.05).Comparing the number of cloned cells among the experimental groups,it was found that there was statistical significance between group 25 ppm and group 75ppm(P <0.05).The results of CCK8 experiment showed that there was statistical significance in cell viability between 25 ppm group and control group after 24 hours of culture(P < 0.05),and there was no statistical significance between75 ppm group,125 pppm group and control group(P > 0.05).After 48 hours of culture,there was no statistical significance in cell viability between experimental groups and control group(P > 0.05).The cell viability in 25 ppm and 75 ppm group was higher than that in control group after 72-and 96-hours culture(P > 0.05).There was no statistical significance in cell viability between the 125 ppm group and the control group(P > 0.05).2.Transwell migration test showed that compared with the control group,the number of cells migrated from the upper chamber to the lower chamber in the experimental groups treated with DDW(125ppm,75 ppm,25ppm)was significantly lower than that in the control group(P < 0.05).At the same time,with the decrease of deuterium concentration in DDW,and the number of cells entering the lower chamber decreased in a dose-dependent manner(P < 0.05).The results of scratch repair test showed that compared with the control group,the area of cell healing in the DDW intervention group(125ppm,75 ppm and 25ppm)was significantly smaller than that in the control group(P< 0.05),and the healing area decreased gradually with the decrease of deuterium concentration in DDW in a dose-dependent manner(P < 0.05).3.Transwell invasion test showed that compared with the control group,the number of cells migrating from the upper chamber to the lower chamber in the experimental groups treated with DDW(125ppm,75 ppm,25ppm)was significantly lower than that in the control group(P < 0.05).At the same time,with the decrease of deuterium concentration in DDW,and the number of cells entering the lower chamber gradually decreased in a dose-dependent manner(P < 0.05).4.The results of Western blot assay showed that the protein contents of MMP-9 and VIM in SKOV3 cells in different concentrations(125ppm,75 ppm,25ppm)of DDW decreased in different degrees,which were significantly lower than those in the control group(P < 0.05)The protein content decreased continuously in a dose-dependent manner with the decrease of deuterium concentration(P < 0.05).Conclusions:1.DDW significantly inhibited the migration and invasion of SKOV3 cells in a dose-dependent manner.2.The mechanism of DDW inhibiting the migration and invasion of ovarian cancer SKOV3 cells may be related to down-regulating the expression of MMP-9 and VIM,and reversing the process of EMT.3.The effect of DDW on the proliferation of ovarian cancer4.SKOV3 cells is not clear,which needs to be further studied.