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Objective: In vitro experiments on phosphate sodium (Creatine Phosphate Sodium, CP) skov3 on human ovarian cancer cell migration and invasion resulting effect. Observed phosphate sodium on skov3 cells invasion and migration affect the results, explore phosphate sodium affect tumor cell invasion and migration possible mechanism for the application of the cardiac toxicity of chemotherapy drugs to provide experimental basis. Methods: Cultured human ovarian cancer cell lines skov3, divided into control group and different concentrations of the drug treatment group. Take good condition logarithmic growth phase cells, using tetrazolium blue assay (MTT assay) detection of phosphate sodium on human ovarian cancer cell adhesion ability of skov3; Transwell chamber assay using phosphoric acid muscle sodium migration ability of the cells and invasion of influence; using flow cytometry phosphate sodium these cell lines nm23-h1, c-myc, MMP-2, MMP-9 protein expression levels of ; using reverse transcription - polymerase chain reaction (revers transcription PCR, RT-PCR) phosphate sodium semi-quantitative detection of the cell lines nm23-h1, c-myc mRNA expression levels. Results: MTT colorimetric analysis testing adhesion experiments showed that: skov3 cell line drug treatment group (1,6,12 mmol / L) compared with the control group, treated with low concentrations of drugs OD values ??did not change significantly, the result was not statistically difference (P gt; 0.05), medium and high concentrations of drug-treated group OD values ??were decreased significantly, the results were significantly different (P lt; 0.05), however, the high concentration of OD values ??between the drug-treated group had no significant change, showed no significant difference (P gt; 0.05); adherence inhibition rates were calculated 0%, 0.72% ± 0.92%, 36.47% ± 3.17%, 37.06% ± 3.30%, treated with low concentrations of drugs inhibiting cell adhesion rate than the control group had no significant change, showed no significant difference (P gt; 0.05), medium and high concentrations of drug-treated group, cell adhesion inhibition rate was significantly higher than the control group, the results were significantly different (P lt; 0.05 ), but no significant change between the two groups, showed no significant difference (P gt; 0.05); namely, a high concentration of drug-treated group skov3 cells with the extracellular matrix components adhesion of Matrigel were significantly inhibited. Invasion assay Show: skov3 cell line drug treatment group (1,6,12 mmol / L) compared with the control group, treated with low concentrations of drugs penetrated cells than the control group did not change significantly, showed no significant difference (P gt ; 0.05), medium and high concentrations of drug-treated group, the number of trans-membrane cells than the control group were significantly reduced, the results were significantly different (P lt; 0.05), however, the high concentration of drug-treated group, the number of cells without penetrating between significant change was not statistically significant (P gt; 0.05); invasion inhibition rates were calculated 0%, 3.06% ± 4.84%, 34.87% ± 3.19%, 37.67% ± 6.06%; namely, a high concentration of drug-treated group on skov3 invasion ability of cells was significantly inhibited. Migration experiments showed that: skov3 cell line drug treatment group (1,6,12 mmol / L) compared with the control group, treated with low concentrations of drugs penetrated cells than the control group did not change significantly, showed no significant difference (P gt ; 0.05), medium and high concentrations of drug-treated group, the number of trans-membrane cells than the control group were significantly reduced, the results were significantly different (P lt; 0.05), however, the high concentration of drug-treated group, the number of cells without penetrating between significant change was not statistically significant (P gt; 0.05); calculating migration inhibition rates were 0%, 2.32% ± 1.47%, 31.50% ± 2.23%, 33.51% ± 2.90%; namely, a high concentration of drug-treated group on skov3 cells ability to migrate significantly inhibited. Protein expression measured by flow cytometry: skov3 cell line drug treatment groups (1,6,12 mmol / L) using the results of flow cytometry, fluorescence index (FI) indicates the expression of four proteins relative content. All test samples are 0mmol / L was the control group, FI of each sample was calculated. FI of nm23-H1 were 1,1.00 ± 0.07,1.27 ± 0.07,1.28 ± 0.05; c-myc for FI values ??were 1,1.00 ± 0.04,0.87 ± 0.01,0.85 ± 0.03; MMP-2 FI of were 1,0.99 ± 0.04,0.98 ± 0.06,0.98 ± 0.03; MMP-9 in FI values ??were 1,1.01 ± 0.03,0.99 ± 0.03,0.98 ± 0.05; namely, a high concentration of drug-treated group skov3 cells nm23 -h1 protein levels compared with the control group was significantly higher, the results were significantly different (P lt; 0.05); medium and high concentrations of drug-treated group skov3 cell line c-myc protein expression levels compared with the control group significantly lower than the results were significantly different (P lt; 0.05); various concentrations of drug-treated group, but skov3 cells MMP-2, MMP-9 protein levels compared with the control group did not change significantly, the result was not statistically Learn the difference (P gt; 0.05). Semi-quantitative RT-PCR analysis showed that: skov3 cell line drug treatment group (1,6,12 mmol / L) using semi-quantitative RT-PCR detection of nm23-h1, c-myc mRNA expression levels of the drug treatment group and the control group both obvious and clearly visible specific bands. Application U.S. FOTODYNE gel imaging analysis system showed that reflect nm23-H1 mRNA expression levels nm23-H1/GAPDH values ??were 0.41 ± 0.02,0.42 ± 0.02,0.63 ± 0.04,0.65 ± 0.03, reflecting the C-myc mRNA expression levels The C-myc/GAPDH values ??were 0.63 ± 0.02,0.62 ± 0.03,0.40 ± 0.01,0.39 ± 0.03; namely, a high concentration of drug-treated group skov3 cells nm23-h1 mRNA expression levels compared with the control group was significantly increases, the results were significantly different (P lt; 0.05); medium and high concentrations of drug-treated group skov3 cell line c-myc mRNA expression levels compared with the control group decreased significantly, the results were significantly different (P lt; 0.05). Conclusion: a certain concentration of phosphate sodium inhibits human ovarian cancer cell skov3 adhesion, migration, invasion role. 2 a certain concentration of phosphate sodium can increase nm23-h1 mRNA and protein levels in human ovarian cancer cell lines skov3 inhibiting adhesion, migration and invasion. 3 a certain concentration of phosphate sodium can lower c-myc mRNA and protein levels suppress human ovarian cancer cell skov3 adhesion, migration and invasion. 4 phosphate sodium for MMP-2, MMP-9 protein expression levels were not significantly affected. 5 phosphate sodium skov3 ovarian cancer cell migration and invasion no role in promoting the application of the drug for the treatment of cardiac toxicity of chemotherapy provides an experimental basis.
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