目的 观察槐耳颗粒联合树突细胞（DC）-细胞因子诱导的杀伤细胞（CIK）对乳腺癌MDA-MB-231干细胞体内外的杀伤效果。方法 用槐耳颗粒（500 mg/L）、槐耳颗粒（1 000 mg/L）与乳腺癌MDA-MB-231细胞作用后，荧光倒置显微镜观察MDA-MB-231细胞形态的变化，MTT比色法检测槐耳颗粒对MDA-MB-231细胞增殖的抑制情况，流式细胞仪检测乳腺癌MDA-MB-231细胞干细胞（CD44⁺CD24^-细胞）所占的比例。采用乳酸脱氢酶（LDH）释放测定法检测DC-CIK细胞联合槐耳颗粒对乳腺癌MDA-MB-231细胞的杀伤作用。建立BalB/C荷MDA-MB-231乳腺癌干细胞裸鼠模型，随机分为模型组、槐耳颗粒组、DC-CIK组及槐耳颗粒与DC-CIK细胞连合组，槐耳颗粒组小鼠按0.33 g/kg ig给药，每天1次，连续3周；DC-CIK组小鼠尾iv DC-CIK细胞5×10⁵个/g，2次/周，连续3周；联合治疗组同时按照单药给药方法联合治疗，观察各组荷瘤裸鼠生存状态，比较各组裸鼠体质量和瘤体积。结果 槐耳颗粒（500 mg/L）、槐耳颗粒（1 000 mg/L）作用于乳腺癌MDA-MB-231细胞36 h后，可使乳腺癌MDA-MB-231细胞逐渐变小，核浓缩，抑制乳腺癌MDA-MB-231细胞的增殖，同时乳腺癌MDA-MB-231细胞中CD44⁺CD24^-细胞比例明显降低。给予荷瘤裸鼠治疗结束后，联合治疗组的瘤质量明显低于2个单独治疗组。结论 槐耳颗粒联合DC-CIK细胞对乳腺癌MDA-MB-231干细胞体内外杀伤效果均好于单独使组。

Objective To study the killing effect of dendritic cells (DC cells)-cytokine-induced killer cells (CIK cells) combined with Huaier Granules on breast cancer MDA-MB-231 stem cells in vivo and in vitro. Methods After the action of the 500 mg/L and the 1 000 mg/L Huaier Granules on breast cancer MDA-MB-231 cells, the morphological changes of MDA-MB-231 cells were observed by fluorescence inverted microscope. The inhibition of the proliferation of MDA-MB-231 cells was detected by MTT colorimetry, and the proportion of MDA-MB-231 cell stem cells (CD44⁺CD24^- cells) in breast cancer was detected by flow cytometry. Lactate dehydrogenase (LDH) release assay was used to detect the killing effect of DC-CIK cells combined with Huaier Granules on breast cancer MDA-MB-231 cells. A nude mouse model of BalB/C bearing MDA-MB-231 breast cancer stem cells was established and randomly divided into four groups: model group, Huaier Granules group, DC-CIK group and the combination group of DC-CIK and Huaier Granules. The mice in the Huaier Granules group were administrated with Huaier Granules by ig at 0.33 g/kg, one time a day for 3 weeks. In DC-CIK group, DC-CIK cells were intravenously injected into the tail of mice 1×10⁵/g, twice a week for 3 weeks; The survival status of tumor bearing nude mice in each group was observed, and the body weight and tumor volume of nude mice in each group were compared. Results After Huaier Granules (500 mg/L) and Huai Er granule (1 000 mg/L) treated on breast cancer MDA-MB-231 cells for 36 h, breast cancer MDA-MB-231 cells were gradually reduced, nuclear concentration, proliferation of breast cancer MDA-MB-231 cells were inhibited, and the proportion of CD44⁺CD24^- cells in MDA-MB-231 cells of breast cancer were significantly reduced. After treatment, the tumor weight of nude mice bearing MDA-MB-231 breast cancer stem cells in the combined treatment group were significantly lower than that of the two separate treatment groups. Conclusion The effect of the combination of DC-CIK and MDA-MB-231 on the breast cancer stem cells in vitro and in vivo is better than that in the separate group.

R969.2

天津市卫生健康委、天津市中医药管理局中、西医结合科研课题（2019035）