二氯乙烷及氯乙酸对细胞周期及凋亡的影响研究

Study on effects of dichloroethane and chloroacetic acid on cell cycle and apoptosis

目的初步探索1,2-二氯乙烷和氯乙酸对大鼠神经胶质瘤细胞和大鼠雪旺细胞周期和凋亡的影响。方法0、15、30、60 mmol/L 1,2-二氯乙烷和0、225、450、900μmol/L氯乙酸分别与大鼠神经胶质瘤细胞和大鼠雪旺细胞共培养24 h,通过流式细胞仪检测细胞周期及凋亡的变化,评价1,2-二氯乙烷和氯乙酸对两种细胞的毒性作用。结果大鼠神经胶质瘤细胞和大鼠雪旺细胞的相对存活率均随着1,2-二氯乙烷和氯乙酸染毒剂量的增加而逐渐降低,1,2-DCE对大鼠神经胶质瘤细胞和大鼠雪旺细胞的IC50值分别为(77.1±1.4)和(72.9±4.3)mmol/L,氯乙酸对大鼠神经胶质瘤细胞和大鼠雪旺细胞的IC50值分别为(895.0±44.5)和(803.2±37.3)μmol/L。0、15、30、60 mmol/L 1,2-二氯乙烷对大鼠神经胶质瘤细胞凋亡率分别为(5.55±0.78)%、(4.93±0.08)%、(6.31±0.53)%和(38.93±2.53)%,G0/G1期细胞比值分别为(61.46±0.79)%、(62.43±2.13)%、(60.19±6.41)%和(43.39±4.82)%,S期细胞比值分别为(13.46±4.57)%、(16.89±3.25)%、(18.56±4.68)%和(32.23±3.49)%,G2/M期细胞比值分别为(23.36±4.53)%、(18.58±5.57)%、(20.14±6.40)%和(23.12±6.46)%。60 mmol/L 1,2-二氯乙烷可引起大鼠神经胶质瘤细胞的凋亡率升高、G0/G1期细胞比值降低、S期细胞比值升高(均P<0.01)。0、15、30、60 mmol/L 1,2-二氯乙烷对大鼠雪旺细胞凋亡率分别为(5.50±1.89)%、(7.95±2.38)%、(7.89±0.99)%和(29.20±7.53)%,G0/G1期细胞比值分别为(39.08±3.20)%、(44.00±20.65)%、(42.15±18.03)%和(37.65±6.39)%,S期细胞比值分别为(28.15±3.62)%、(25.77±15.47)%、(25.20±7.58)%和(23.85±2.00)%,G2/M期细胞比值分别为(32.02±2.78)%、(29.58±7.06)%、(31.49±12.27)%和(37.10±7.68)%。60 mmol/L 1,2-二氯乙烷可引起大鼠雪旺细胞凋亡率升高(P<0.01)。0、225、450、900μmol/L氯乙酸对大鼠神经胶质瘤细胞凋亡率分别为(5.55±0.78)%、(6.87±4.07)%、(5.16±0.95)%和(5.70±1.21)%,G0/G1期细胞比值分别为(61.46±0.79)%、(60.15±7.92)%、(61.27±3.11)%和(63.57±3.51)%,S期细胞比值分别为(13.46±4.57)%、(18.81±5.95)%、(16.92±4.68)%和(13.99±1.34)%,G2/M期细胞比值分别为(23.36±4.53)%、(18.93±1.96)%、(19.90±4.91)%和(20.86±2.03)%。0、225、450、900μmol/L氯乙酸对大鼠雪旺细胞凋亡率分别为(5.50±1.89)%、(7.60±2.60)%、(9.20±2.92)%和(37.88±6.99)%,G0/G1期细胞比值分别为(39.08±3.20)%、(39.88±6.06)%、(37.30±9.58)%和(36.59±7.16)%,S期细胞比值分别为(28.15±3.62)%、(20.91±7.76)%、(22.01±7.28)%和(16.72±5.49)%,G2/M期细胞比值分别为(32.02±2.78)%、(38.54±2.71)%、(38.74±5.21)%和(45.49±5.70)%。900μmol/L氯乙酸可引起大鼠雪旺细胞凋亡率和G2/M期升高(均P<0.05)。结论1,2-二氯乙烷可引起大鼠神经胶质瘤细胞和大鼠雪旺细胞凋亡率升高、大鼠神经胶质瘤细胞周期阻滞。氯乙酸可引起大鼠雪旺细胞凋亡率升高、细胞周期阻滞。

Objective To preliminarily explore the effects of 1,2-dichloroethane and chloroacetic acid on the cell cycle and apoptosis in rat glioma cells and rat Schwann cells.Methods 0,15,30,60 mmol/L 1,2-dichloroethane and 0,225,450,900μmol/L chloroacetic acid were co-cultured with rat glioma cells and rat Schwann cells for 24 h,respectively.The changes in the cell cycle and apoptosis were detected by flow cytometry,to evaluate the toxicity of 1,2-dichloroethane and chloroacetic acid in the two cells.Results With the increasing toxic doses of 1,2-dichloroethane and chloroacetic acid,the relative cell survival rate was gradually decreased in both cells.The IC50 value of 1,2-dichloroethane was(77.1±1.4)and(72.9±4.3)mmol/L in rat glioma cells and rat Schwann cells respectively.The IC50 value of chloroacetic acid was(895.0±44.5)and(803.2±37.3)μmol/L in rat glioma cells and rat Schwann cells respectively.The apoptosis rates of 0,15,30,60 mmol/L 1,2-dichloroethane in rat glioma cells were respectively(5.55±0.78)%,(4.93±0.08)%,(6.31±0.53)%and(38.93±2.53)%,the cell ratios in G0/G1 phase were respectively(61.46±0.79)%,(62.43±2.13)%,(60.19±6.41)%and(43.39±4.82)%,in S phase were respectively(13.46±4.57)%,(16.89±3.25)%,(18.56±4.68)%and(32.23±3.49)%,in G2/M phase were respectively(23.36±4.53)%,(18.58±5.57)%,(20.14±6.40)%and(23.12±6.46)%.1,2-dichloroethane can cause an increased rate of apoptosis,lower cell ratio in G0/G1 phase and increased cell ratio in S phase in rat glioma cells at the dose of 60 mmol/L(all P<0.01).The apoptosis rates of 0,15,30,60 mmol/L 1,2-dichloroethane in rat Schwann cells were respectively(5.50±1.89)%,(7.95±2.38)%,(7.89±0.99)%and(29.20±7.53)%,and the cell ratios in G0/G1 phase were respectively(39.08±3.20)%,(44.00±20.65)%,(42.15±18.03)%and(37.65±6.39)%,in S phase were respectively(28.15±3.62)%,(25.77±15.47)%,(25.20±7.58)%and(23.85±2.00)%,in G2/M phase were respectively(32.02±2.78)%,(29.58±7.06)%,(31.49±12.27)%and(37.10±7.68)%.60 mmol/L 1,2-dichloroethane can cause an increased rate of apoptosis in rat Schwann cells(P<0.01).The apoptosis rates of 0,225,450,900μmol/L chloroacetic acid in rat glioma cells were respectively(5.55±0.78)%,(6.87±4.07)%,(5.16±0.95)%and(5.70±1.21)%,and the cell ratios in G0/G1 phase were respectively(61.46±0.79)%,(60.15±7.92)%,(61.27±3.11)%and(63.57±3.51)%,in S phase were respectively(13.46±4.57)%,(18.81±5.95)%,(16.92±4.68)%and(13.99±1.34)%,in G2/M phase were respectively(23.36±4.53)%,(18.93±1.96)%,(19.90±4.91)%and(20.86±2.03)%.The apoptosis rates of 0,225,450,900μmol/L chloroacetic acid in rat Schwann cells were respectively(5.50±1.89)%,(7.60±2.60)%,(9.20±2.92)%and(37.88±6.99)%,and the cell ratios in G0/G1 phase were respectively(39.08±3.20)%,(39.88±6.06)%,(37.30±9.58)%and(36.59±7.16)%,in S phase were respectively(28.15±3.62)%,(20.91±7.76)%,(22.01±7.28)%and(16.72±5.49)%,in G2/M phase were respectively(32.02±2.78)%,(38.54±2.71)%,(38.74±5.21)%and(45.49±5.70)%.900μmol/L chloroacetic acid can cause an increased rate of apoptosis and increased cell ratio in G2/M phase in rat Schwann cells(both P<0.05).Conclusion 1,2-dichloroethane can cause increased apoptotic rate of rat glioma cells and Schwann cells,cycle arrest in rat glioma cells.Chloroacetic acid can cause increased apoptotic rate and cycle arrest in rat Schwann cells.