目的 觀察槐耳顆粒聯(lián)合樹突細(xì)胞（DC）-細(xì)胞因子誘導(dǎo)的殺傷細(xì)胞（CIK）對(duì)乳腺癌MDA-MB-231干細(xì)胞體內(nèi)外的殺傷效果。方法 用槐耳顆粒（500 mg/L）、槐耳顆粒（1 000 mg/L）與乳腺癌MDA-MB-231細(xì)胞作用后，熒光倒置顯微鏡觀察MDA-MB-231細(xì)胞形態(tài)的變化，MTT比色法檢測(cè)槐耳顆粒對(duì)MDA-MB-231細(xì)胞增殖的抑制情況，流式細(xì)胞儀檢測(cè)乳腺癌MDA-MB-231細(xì)胞干細(xì)胞（CD44⁺CD24^-細(xì)胞）所占的比例。采用乳酸脫氫酶（LDH）釋放測(cè)定法檢測(cè)DC-CIK細(xì)胞聯(lián)合槐耳顆粒對(duì)乳腺癌MDA-MB-231細(xì)胞的殺傷作用。建立BalB/C荷MDA-MB-231乳腺癌干細(xì)胞裸鼠模型，隨機(jī)分為模型組、槐耳顆粒組、DC-CIK組及槐耳顆粒與DC-CIK細(xì)胞連合組，槐耳顆粒組小鼠按0.33 g/kg ig給藥，每天1次，連續(xù)3周；DC-CIK組小鼠尾iv DC-CIK細(xì)胞5×10⁵個(gè)/g，2次/周，連續(xù)3周；聯(lián)合治療組同時(shí)按照單藥給藥方法聯(lián)合治療，觀察各組荷瘤裸鼠生存狀態(tài)，比較各組裸鼠體質(zhì)量和瘤體積。結(jié)果 槐耳顆粒（500 mg/L）、槐耳顆粒（1 000 mg/L）作用于乳腺癌MDA-MB-231細(xì)胞36 h后，可使乳腺癌MDA-MB-231細(xì)胞逐漸變小，核濃縮，抑制乳腺癌MDA-MB-231細(xì)胞的增殖，同時(shí)乳腺癌MDA-MB-231細(xì)胞中CD44⁺CD24^-細(xì)胞比例明顯降低。給予荷瘤裸鼠治療結(jié)束后，聯(lián)合治療組的瘤質(zhì)量明顯低于2個(gè)單獨(dú)治療組。結(jié)論 槐耳顆粒聯(lián)合DC-CIK細(xì)胞對(duì)乳腺癌MDA-MB-231干細(xì)胞體內(nèi)外殺傷效果均好于單獨(dú)使組。

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

天津市衛(wèi)生健康委、天津市中醫(yī)藥管理局中、西醫(yī)結(jié)合科研課題（2019035）