顺铂联合加热对体外血液中肝肿瘤细胞的杀灭作用及对红细胞的影响

目的 探讨顺铂(DDP)联合加热对体外血液中肝肿瘤细胞的杀灭作用及对红细胞的影响.方法 将肝肿瘤细胞2 ml加入10 ml红细胞悬液中,充分混匀后,分别取2 ml于离心管中.采用随机数字表法,将其随机分为6组(n=30):A组、D组各加入2 ml生理盐水,B组、E组各加入2 mlDDP(终浓度为100 μg/ml),C组和F组各加入2 ml DDP(终浓度为200 μg/ml).充分混匀后,A组、B组和C组放入37℃恒温水浴箱中,其余3组放入42℃恒温水浴箱中进行加热处理,处理时间均为30min.采用密度梯度离心法分离肝肿瘤细胞和红细胞,测定肝肿瘤细胞抑制率及克隆形成率,测定红细胞渗透脆性及2,3-二磷酸甘油酸(2,3-DPG)含量.结果 随温度和DDP浓度的升高,肿瘤细胞的抑制率升高,克隆形成率降低(P<0.01).F组肿瘤细胞抑制率达98%以上,且未出现克隆形成.处理前后F组红细胞2,3-DPG含量比较差异无统计学意义(P->0.05).处理后F组红细胞在0.68%NaCl溶液中的溶血率<1%.结论 200 μg/ml DDP联合42 ℃加热30 min,可使体外血液中肝肿瘤细胞失去增殖能力,而对红细胞膜的影响轻微,对红细胞的携氧能力无影响.

Abstract：

Objective To investigate the effectiveness of cis-diamminedichloroplatinum (DDP) combined with hyperthennia in killing liver tumor cells and its influence on erythrocytes in vitro. Methods Cultured liver tumor cells (2 ml) were mixed with erythrocyte suspension (10 ml) and then the mixture was separated into 6 centrifuge tubes with 2 ml in each one. The centrifuge tubes were randomly divided into A-F groups and the experiment was repeated for 30 times. Normal saline 2 ml was added in A and D groups. DDP 2 ml (200 μg/ml) was added in B and E groups. DDP 2 ml (400 μg/ml) was added in C and F groups. The cells were then incubated in warm bath of 37 ℃ for 30 min in A, B and C groups and in warm bath of 42 ℃ for 30 min in the other three groups.After hyperthermic treatment, tumor cells were isolated from erythrocytes using density gradient centrifugation, the inhibition rate of tumor cells was determined by MTT method and the clone formation of tumor cells was checked.The erythrocyte osmotic fragility and content of 2,3-diphosphoglyceric acid in erythrocytes were measured. Results The inhibition rate of tumor cells was gradually increased, while the rate of tumor cell clone formation decreased with the increase in the temperature and DDP concentrations ( P < 0.01) . The rate of tumor cell clone formation was more than 98% and no clone formation was tested in group F. There was no significant difference in the content of 2,3-diphosphoglyceric acid in erythrocytes between before and after hyperthermic treatment in group F ( P >0.05 ) . The rate of hemolysis of erythrocytes was less than 1 % in the 0.68 % sodium chloride solution in group F.Conclusion DDP 200 μg/ml combined with hyperthermic treatment with temperature of 42 ℃ for 30 min can make the liver tumor cells lose the capability of proliferation, however, it exerts slight effect on erythrocyte membrane and no influence on the oxygen-carrying capacity of erythrocytes.

Effectiveness of cis-diamminedichloroplatinum combined with hyperthermia in killing liver tumor cells and its influence on erythrocytes in vitro

Objective To investigate the effectiveness of cis-diamminedichloroplatinum (DDP) combined with hyperthennia in killing liver tumor cells and its influence on erythrocytes in vitro. Methods Cultured liver tumor cells (2 ml) were mixed with erythrocyte suspension (10 ml) and then the mixture was separated into 6 centrifuge tubes with 2 ml in each one. The centrifuge tubes were randomly divided into A-F groups and the experiment was repeated for 30 times. Normal saline 2 ml was added in A and D groups. DDP 2 ml (200 μg/ml) was added in B and E groups. DDP 2 ml (400 μg/ml) was added in C and F groups. The cells were then incubated in warm bath of 37 ℃ for 30 min in A, B and C groups and in warm bath of 42 ℃ for 30 min in the other three groups.After hyperthermic treatment, tumor cells were isolated from erythrocytes using density gradient centrifugation, the inhibition rate of tumor cells was determined by MTT method and the clone formation of tumor cells was checked.The erythrocyte osmotic fragility and content of 2,3-diphosphoglyceric acid in erythrocytes were measured. Results The inhibition rate of tumor cells was gradually increased, while the rate of tumor cell clone formation decreased with the increase in the temperature and DDP concentrations ( P < 0.01) . The rate of tumor cell clone formation was more than 98% and no clone formation was tested in group F. There was no significant difference in the content of 2,3-diphosphoglyceric acid in erythrocytes between before and after hyperthermic treatment in group F ( P >0.05 ) . The rate of hemolysis of erythrocytes was less than 1 % in the 0.68 % sodium chloride solution in group F.Conclusion DDP 200 μg/ml combined with hyperthermic treatment with temperature of 42 ℃ for 30 min can make the liver tumor cells lose the capability of proliferation, however, it exerts slight effect on erythrocyte membrane and no influence on the oxygen-carrying capacity of erythrocytes.