CIK对地西他滨增敏乳腺癌MDA-MB-231细胞的杀伤机制探讨

目的 观察地西他滨（DAC）增敏细胞因子介导的杀伤细胞（CIK）对人乳腺癌MDA-MB-231细胞株的细胞毒作用,并探讨其增敏机制.方法 取对数生长期的乳腺癌MDA-MB-231及正常乳腺MCF-10A细胞,MMT法检测DAC及TRAIL对乳腺癌细胞的增殖抑制率,LDH法检测DAC单用或联合肿瘤坏死因子相关凋亡诱导配体（TRAIL）及CIK对乳腺癌细胞的杀伤率,流式细胞仪检测细胞凋亡率.结果 DAC对乳腺癌细胞MDA-MB-231及正常乳腺上皮细胞MCF-10A无明显杀伤作用.TRAIL对MDA-MB-231有明显杀伤作用,作用24h的IC50约为100 ng/mL,然而对MCF-10A无明显促凋亡影响.5∶1、10∶1、20∶1效靶比的CIK对MDA-MB-231的杀伤率分别为10.7％±1.2％、17.8％±2.1％、37.2％±3.4％,对MCF-10A无杀伤作用.经50 μmol/L的DAC预处理乳腺癌细胞MDA-MB-231 24 h后,5∶1、10∶1、20∶1效靶比的CIK对MDA-MB-231的杀伤率分别为18.6％±2.6％、26.3％±4.5％、51.6％±6.6％,与相应单独效靶比的CIK杀伤作用比较,P均＜0.01.结论 DAC增敏CIK对乳腺癌MDA-MB-231细胞的杀伤敏感性,对正常的乳腺上皮细胞无影响,DAC联合CIK可能成为治疗乳腺癌患者的新途径.

Decitabine sensitizes breast cancer cells MDA-MB-231 to CIK-mediated cell death

Objective To investigate the cytotoxic effects of decitabine associated with cytokine-induced killer （CIK） ceils against MDA-MB-231 cells of breast cancer and to discusse the possible molecular mechanisms. Methods The breast cancer cells MDA-MB-231 and normal breast cell line MCF-10A at the logarithmic growth phase were selected and MTr assay was used to detect the proliferation inhibition rate treated by DAC and TRAIL. Lactate dehydrogenase （LDH） assay was used to measure the influence on cytotoxic activity of DAC or TRAIL combined with CIK cells against breast canc- er cells in vitro, and the apoptosis rate was detected by flow cytometry. Results DAC had low cytotoxic effects on breast cancer MDA-MB-231 cells and normal breast cell line MCF-10A. TRAIL had obviously cytotoxic effects on MDA-MB-231 ceils and the ICs0 for 24 h was about 100 ng/mL, while had no effect on normal breast cell line MCF-10A. The effect-target of 5： 1, 10：1 and 20：1 CIK would induce 10.7 % ± 1.2%, 17.8 % ± 2.1%, and 37.2%±3.4% apoptosis of MDA-MB- 231 cells, but had no effect on MCF-10A. Pretreatment with 50 μmol/L of DAC for 24 h, the effect-target of 5： 1, 10：1 and 20：1 CIK would induce 18.6%±2.6%, 26.3% ±4.5%, 51.6% ±6.6% apoptosis of MDA-MB-231 cells. When we compared with relevant effect-target of CIK, significant different was found （ all P 〈 0.01 ）. Conclusion DAC could sensitize breast cancer cells MDA-MB-231 to CIK and has no cytotoxic effect on normal breast cell line MCF-10A. There- fore, DAC combined with CIK may be a new way for the treatment of patients with breast cancer.