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《The Journal of Traditional Chinese Orthopedics and Traumatology》 2017-01
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Effect of sodium dichloroacetate on cell proliferation,apoptosis and migration in human MG63 osteosarcoma cells

YU Yali;WANG Leiming;Zhengzhou Orthopedic Hospital;  
Objective: To explore the effect of sodium dichloroacetate( DCA- Na) on cell proliferation,apoptosis and migration in human MG63 osteosarcoma cells. Methods: The MG63 cells were randomly divided into control group,low- concentration DCA- Na group,middle- concentration DCA- Na group and high- concentration DCA- Na group in the logarithmic growth phase. No DCA- Na were placed in cells in control group,and DCA- Na with final concentration of 50,100 and 200 μg / m L were placed in cells in low-,middle- and high- concentration DCA- Na group respectively,meanwhile,a blank group was established with same amount of culture medium and without cells and DCA- Na. At 24,48 and 72 hours after the intervention,the MG63 cell proliferations were detected by using Cell Counting Kit- 8 spectrophotometry and the optical density( OD) were measured for calculating the cell proliferation inhibition rate( [1-( ODDCA- Na group- ODblank group) /( ODcontrol group- ODblank group) ]× 100%) of low-,middle- and high- concentration DCA- Na group respectively. Meanwhile,The Caspase- 3 enzymatic activity and apoptosis of MG63 cells were detected by using Caspase-3 activity assay kit spectrophotometry and Annexin V- FITC apoptosis assays kit flow cytometry respectively. The MG63 cell migration was detected by using Transwell assay at 48 hours after the intervention. Results: The MG63 cell proliferations were obviously inhibited after intervention in low-,middle- and high- concentration DCA- Na group,and a time- dependent rising trend of cell proliferation inhibition rate was found in the 3 groups. There was statistical difference in the MG63 cell proliferation inhibition rate between different timepoints( F = 847. 080,P = 0. 000),in other words,there was time effect. There was statistical difference in the MG63 cell proliferation inhibition rate between low-,middle- and high- concentration DCA- Na group at 24,48 and 72 hours after the intervention,and the MG63 cell proliferation inhibition rate was lower in the low- concentration DCA- Na group compared to middle- concentration DCA- Na group and was lower in the middle- concentration DCA- Na group compared to high- concentration DCA- Na group( 10. 802 +/- 3. 000,18. 792 +/-2. 261,27. 080 + /- 3. 133%,F = 2. 795,P = 0. 000; 13. 098 + /- 1. 299,25. 215 + /- 2. 676,44. 382 + /- 2. 397%,F = 4. 362,P = 0. 000;14. 728 + /- 1. 177,35. 297 + /- 4. 757,64. 227 + /- 4. 549%,F = 5. 896,P = 0. 000). There was statistical difference in MG63 cell proliferation inhibition rate between the 3 groups in general,in other words,there was group effect( F = 296. 412,P = 0. 000). There was interaction between time factor and group factor( F = 75. 678,P = 0. 000). No significant change was found in Caspase- 3 enzymatic activity of MG63 cells in control group,while a rising trend of the Caspase- 3 enzymatic activity of MG63 cells was found in low-,middle- and high- concentration DCA- Na group after intervention. There was statistical difference in the Caspase- 3 enzymatic activity of MG63 cells between different timepoints( F = 1 480. 792,P = 0. 000),in other words,there was time effect. There was statistical difference in the Caspase- 3enzymatic activity of MG63 cells between the 4 groups at 24,48 and 72 hours after the intervention,and the Caspase- 3 enzymatic activity of MG63 cells presented a low- to- high trend in control group and low-,middle- and high- concentration DCA- Na group in turn( 0. 027 +/- 0. 003,0. 143 +/- 0. 005,0. 153 +/- 0. 008,0. 161 +/- 0. 003,F = 2. 320,P = 0. 000; 0. 035 +/- 0. 003,0. 174 +/- 0. 004,0. 184 + /- 0. 007,0. 253 + /- 0. 001,F = 1. 014,P = 0. 000; 0. 031 + /- 0. 004,0. 246 + /- 0. 006,0. 275 + /- 0. 003,0. 371 + /- 0. 004,F =1. 000,P = 0. 000). There was statistical difference in Caspase- 3 enzymatic activity of MG63 cells between the 4 groups in general,in other words,there was group effect( F = 624. 975,P = 0. 000). There was interaction between time factor and group factor( F = 94. 579,P =0. 000). No significant change was found in MG63 cell apoptosis rate in control group,while a rising trend of MG63 cell apoptosis rate was found in low-,middle- and high- concentration DCA- Na group after intervention. There was statistical difference in the MG63 cell apoptosis rate between different timepoints( F = 359. 645,P = 0. 000),in other words,there was time effect. There was statistical difference in the MG63 cell apoptosis rate between the 4 groups at 24,48 and 72 hours after the intervention,and the MG63 cell apoptosis rate presented a low- to- high trend in control group and low-,middle- and high- concentration DCA- Na group in turn( 2. 554 + /- 0. 427,10. 708 + /- 2. 012,20. 857 + /- 2. 531,27. 312 + /- 2. 140%,F = 6. 733,P = 0. 000; 1. 748 + /- 0. 202,18. 604 + /- 2. 721,29. 471 + /-1. 605,36. 873 + /- 2. 734%,F = 9. 292,P = 0. 000; 1. 944 + /- 0. 112,24. 071 + /- 3. 921,30. 050 + /- 3. 921,38. 211 + /- 1. 721%,F =8. 237,P = 0. 000). There was statistical difference in the MG63 cell apoptosis rate between the 4 groups in general,in other words,there was group effect( F = 46. 627,P = 0. 000). There was interaction between time factor and group factor( F = 7. 012,P = 0. 000). There was statistical difference in the number of migratory MG63 cells under the optical microscope( × 200) between the 4 groups at 48 hours after the intervention( 84. 45 + /- 10. 45,74. 56 + /- 9. 45,65. 41 + /- 5. 21,40. 21 + /- 4. 52,F = 148. 243,P = 0. 000). The migratory MG63 cells was less in low-,middle- and high- concentration DCA- Na group compared to control group( P = 0. 017,P = 0. 001,P = 0. 000),and the low- concentration DCA- Na group surpassed middle- and high- concentration DCA- Na group and middle- concentration DCA-Na group surpassed high- concentration DCA- Na group in the number of migratory MG63 cells( P = 0. 012,P = 0. 001,P = 0. 000).Conclusion: DCA- Na can inhibit the proliferation of human MG63 osteosarcoma cells,increase the Caspase- 3 enzymatic activity ofMG63 cells,induce and accelerate the apoptosis of MG63 cells and inhibit the migration of MG63 cells. Moreover,the higher the concentration of DCA- Na is and the longer the intervention time is,the more obvious the effect is.
【CateGory Index】: R738
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