r 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 KG1a proliferation and the synergism could increase apoptosis rate.
Key words: daunorubicin;thermotherapy;acute myeloblastic leukemia;KG1a cell lines;antiproliferation;apoptosis
随着新的化疗药物和靶向治疗药物的出现,白血病的疗效取得了很大的进步。但难治/复发性白血病的完全缓解率低,长期生存率不高,治疗效果仍然不够理想。单纯依靠增加化疗药物剂量势必带来较大的毒副作用,如何提高白血病细胞对化疗药物的敏感性,克服白血病细胞耐药是当前白血病治疗中亟待解决的问题。近年来,随着热疗理论和技术的发展,热疗已成为继手术、化疗、放疗、生物治疗之后的第5大肿瘤治疗手段,热疗与其他治疗方法联合有协同作用[1]。热化疗在实体肿瘤治疗中研究较多[2],但对血液系统恶性疾病研究较少。人急性髓白血病细胞KG1a高表达CD34+抗原,对化疗药物不敏感[3],本研究观察热疗联合柔红霉素(daunorubicin,DNR)对其体外增殖及凋亡的影响,为临床热疗联合化疗治疗复发/难治性白血病提供理论和实验依据。
1 材料与方法
1.1 细胞株
人急性髓细胞白血病细胞株KG1a由中国医学科学院血液学研究所馈赠,由本实验室传代保存。用含体积分数10%胎牛血清的RPMI1640培养液于37 ℃、体积分数5%CO2的培养箱中培养。实验中所用细胞均处于对数生长期。
1.2 试剂
RPMI1640 培养基(Gibco公司);胎牛血清(杭州四季青生物公司);甲基偶氮唑蓝(methyl thiazolyl tetrazolium,MTT)(Sigma公司)使用时用磷酸盐缓冲液(phosphate buffer solution,PBS)配成5 mg·mL-1的溶液[4];DNR(浙江海正药业有限公司);二甲基亚砜(dimethyl sulfoxide,DMSO)(Sigma公司);甲基纤维素(Sigma公司)。
1.3 仪器
恒温水浴箱(上海医用恒业设备厂),精密加样枪(美国Eppendorf公司),ELX800酶标仪(美国BioTek公司),倒置相差显微镜(德国Leica公司),Eppendorf低温高速离心机(美国Fisher Scientific公司),ELITE型流式细胞仪(美国Coulter公司)。
1.4 方法
1
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