热疗联合柔红霉素对人急性髓细胞白血病KG1a细胞增殖抑制及凋亡的影响

Effects of thermotherapy combined with daunorubicin on the anti-proliferation and apoptosis of leukemia cell lines KG1a

目的观察热疗联合柔红霉素(DNR)对人急性髓细胞白血病细胞株KG1a体外增殖抑制和凋亡的影响。方法甲基偶氮唑蓝(MTT)法检测DNR作用KG1a细胞48 h抑制50%的药物浓度(IC50),以此浓度为工作浓度,设对照组(正常培养)、热疗组(单纯加热)、化疗组(单纯DNR处理)和热化疗组(加热联合DNR处理),热疗和热化疗各设40℃、42℃、43℃3个温度组,热疗在恒温水浴箱中加热60 min;MTT法检测作用前后对KG1a细胞的抑制作用;甲基纤维素培养体系分析细胞克隆能力;流式细胞仪检测KG1a细胞凋亡率。结果热疗、化疗和热化疗均能抑制KG1a细胞增殖,热化疗组细胞抑制率明显高于化疗组及相同温度的热疗组,且抑制作用随温度的升高而增强(P<0.05);热疗组、化疗组和热化疗组细胞克隆形成率明显低于对照组(P<0.05),热化疗组克隆集体落形成率明显低于热疗组、化疗组(P<0.05);热疗组、化疗组和热化疗组的细胞凋亡率均较对照组显著升高(P<0.05),热化疗组与化疗组及相同温度热疗组相比细胞凋亡率均显著提高(P<0.05)。结论热疗联合DNR能增强对KG1a细胞的体外抑制作用,提高其凋亡率。

Objective To observe the effects of thermotherapy combining with daunorubicin（DNR） on the anti-proliferation and apoptosis of leukemia cell lines KG1 a. Methods IC50 of DNR against KG1 a cell was measured by MTT assay after treated for 48 hours, and took it as the working concentration in following experiment. Found control group （normal culturing）, thermotherapy group（ heating alone ）, chemotherapy group （ treating by DNR only ）, thermotherapy combining with chemotherapy group（ beating combining with DNR）. Thermotherapy group and thermotherapy combining with chemotherapy group found three subgroup respectively, such as 40 ℃ ,42 ℃ ,43 ℃ temperature group. Thermotherapy group was heated in the thermostatic waterbath for 60 min. The inhibition rate of proliferation was evaluated by MTT assay and clonality was observed in cellosolve system, the rate of apoptosis was analyzed by flow cytometry. Results Thermotherapy group,chemotherapy group,tbermotherapy combining with chemotherapy group could inhibit KG1 a cell proliferation, the inhibition ratio in thermotherapy combining with chemotherapy group was higher than that in chemotherapy group and the thermotherapy group in the same temperature, and the effect of inhibition enhanced with the temperature rising（ P 〈 0. 05 ）. The inhibition was enhanced when temperature increasing. Compared with control group, clonality of Thermotherapy, chemotherapy, thermotherapy combining with chemotherapy group was lower, but the rate of apoptosis of them was higher（ P 〈 0.05 ）. Compared with thermotherapy group and chemotherapy group, the difference of clonality of thermotherapy combining with chemotherapy group has statistical significance （ P 〈 0.05 ）. Also the difference of the rate of apoptosis of thermotherapy combining with chemotherapy group has statistical significance compared chemotherapy group and thermotherapy group in the same temperature （ P 〈 0.05 ）. Conclusion Thermotherapy combined with DNR could obviously inhibit KG1 a proliferation and the synergism could increase apoptosis rate.