Study on the mechanism of curcumin on the apoptosis and radiosensitivity of human chordoma cell line CM-319 / 中华骨科杂志

ABSTRACT

Objective:To investigate the effects of curcumin on apoptosis and radiosensitivity of human chordoma cell line CM-319 and its mechanism.Methods:CM-319 cells were cultured in vitro and divided into curcumin 0 μmol/L group, curcumin 5 μmol/L group, curcumin 10 μmol/L group, curcumin 20 μmol/L group, si-NC group, si-UHRF1 group, curcumin+pcDNA group, curcumin+pcDNA-UHRF1 group.Flow cytometry was usedtodetect cell apoptosis, Western Blot was used to detectthe protein expression of Bcl-2, Bax, Cleaved Caspase-3 and UHRF1, qRT-PCR was used to detect the expression of UHRF1 mRNA, and clone formationexperiment was used to detect the sensitivity of CM-319 cells to radiation.Results:Compared with the curcumin 0 μmol/L group, the apoptosis rate of CM-319 cellsinthe curcumin 5 μmol/L group, curcumin 10 μmol/L group and curcumin 20 μmol/L groupincreased 12.61%±1.57%, 18.42%±1.54%, 29.37%±2.65% vs 7.24%±0.73% ( t=9.305, 19.680, 24.153; all P<0.05), and the protein expression of Bcl-2 decreased 0.62±0.06, 0.51±0.05, 0.33±0.03 vs 0.76±0.07 ( t=4.556, 8.719, 16.939; all P<0.05), while the protein expression of Bax 0.41±0.04, 0.55±0.05, 0.69±0.06 vs 0.26±0.03 ( t=9.000, 14.920, 19.230; all P<0.05) and Cleaved Caspase-3 0.37±0.04, 0.49±0.05, 0.62±0.06 vs 0.24±0.03 ( t=7.800, 12.862, 16.994; all P<0.05) increased, and the expression of UHRF1 mRNA decreased 0.82±0.08, 0.67±0.07, 0.42±0.04 vs 1.01±0.09 ( t=4.734, 8.946, 17.972; all P<0.05), the protein expression of UHRF1 decreased 0.61±0.06, 0.48±0.05, 0.31±0.03 vs 0.83±0.08 ( t=6.600, 11.130, 18.258; all P<0.05), and were dose-dependent, and the sensitization ratios were 1.433, 1.708, and 2.183, respectively. Compared with the si-NC group, the protein expression of UHRF1in CM-319 cells in the si-UHRF1 group decreased 0.35±0.04 vs 0.79±0.08 ( t=14.758, P<0.05), the apoptosis rate of CM-319 cells increased 21.49%±2.11% vs 8.24%±0.83% ( t=17.531, P<0.05), the protein expression of Bcl-2 decreased 0.34±0.03 vs 0.71±0.07 ( t=14.575, P<0.05), while the protein expression of Bax 0.62±0.06 vs 0.25±0.03 ( t=16.547, P<0.05) and Cleaved Caspase-3 0.58±0.05 vs 0.23±0.03 ( t=18.007, P<0.05) increased, and the sensitization ratio was 1.727. Compared with the curcumin+pcDNA group, the apoptosis rate of CM-319 cells in the curcumin+pcDNA-UHRF1 groupreduced 13.59%±1.25% vs 28.31%±3.01% ( t=13.549, P<0.05), the protein expression of Bcl-2 increased 0.63±0.06 vs 0.31±0.03 ( t=14.311, P<0.05), while the protein expression of Bax 0.42±0.04 vs 0.71±0.07 ( t=10.791, P<0.05) and Cleaved Caspase-3 0.38±0.04 vs 0.65±0.05 ( t=12.650, P<0.05) decreased, and the sensitization ratio was 0.539. Conclusion:Curcumin can induce apoptosis of CM-319 cells and enhance the radiosensitivity of CM-319 cells, , and the mechanism is related to the down-regulation of UHRF1 expression.