不同周期新辅助化疗后乳腺癌干细胞的分离培养和体外增殖特性

Isolation, culture and in vitro proliferation of breast cancer stem cells after different cycles of neoadjuvant chemotherapy

背景:接受不同周期新辅助化疗的乳腺癌组织中是否都存在乳腺癌化疗微球体以及其对乳腺癌干细胞分离培养的影响尚不清楚。目的:探讨不同周期新辅助化疗后乳腺癌组织乳腺癌干细胞的体外增殖特性。方法:从不同周期新辅助化疗后的乳腺癌组织中分离微球体,绘制细胞生长曲线,免疫细胞化学检测乳腺癌干细胞标记物ALDH1表达情况。结果与结论:从新辅助化疗1个周期的标本中无法获得微球体,化疗2,3,4个周期的部分标本中可以培养出微球体。在培养的前3 d,化疗3个周期和化疗2个周期细胞数差异无显著性意义,但在3 d之后,化疗3个周期的细胞增殖速度显著快于化疗2个周期,6 d之后,化疗2个周期的细胞生长曲线呈现出平缓的状态,化疗3个周期则呈现出迅速上升的状态。化疗3个周期标本的乳腺癌干细胞微球体中ALDH1阳性细胞比例大于化疗2个周期。结果表明化疗2、3个周期的乳腺癌干细胞均具有一定的增殖和分化能力,从化疗后的乳腺癌组织中分离乳腺癌干细胞宜选取化疗3个周期及以上的标本。

BACKGROUND： Whether there are breast cancer stem cell microspheres in the breast cancer tissues and whether these microspheres have an impact on isolation and culture of breast cancer stem cells after different cycles of neoadjuvant chemotherapy are still unclear. OBJECTIVE： To explore the proliferation and differentiation of breast cancer stem cells in breast cancer tissues after different cycles of neoadjuvant chemotherapy. METHODS： Breast cancer stem cell microspheres were isolated from the breast cancer tissues after different cycles of neoadjuvant chemotherapy to drawn a cell growth curve. Immunocytochemical method was used to detect ALDH1 expression. RESULTS AND CONCLUSION： Microspheres could be obtained from the specimens of neoadjuvant chemotherapy for two, three and four cycles rather than one cycle. At 3 days prior to culture, there was no difference in the number of cells isolated after two- and three-cycle neoadjuvant chemotherapy; but after 3 days, the cells from the three-cycle neoadjuvant chemotherapy proliferated faster than those from the two-cycle neoadjuvant chemotherapy; after 6 days, the cell growth curve of two-cycle neoadjuvant chemotherapy was in the plateau stage, and the proliferation of cells from the three-cycle neoadjuvant chemotherapy showed a rapid increase trend. The positive expression of ALDH1 in the microspheres from the three-cycle neoadjuvant chemotherapy was higher than that from the two-cycle neoadjuvant chemotherapy. These findings indicate that breast cancer stem cells from the specimens of two- and three-cycle neoadjuvant chemotherapy both have proliferation and differentiation potentials, and the specimens of three-cycle neoadjuvant chemotherapy or above are preferred.