去胰岛素的乳腺癌干细胞培养及雌激素诱导的作用

 目的：探讨去除胰岛素的乳腺癌干细胞悬浮培养方法以及雌激素诱导的作用。方法：通过使用去除胰岛素的悬浮培养方法获得乳腺癌细胞系MCF7的肿瘤球细胞，并通过形态学观察、CD24-CD44+表达细胞的检测、醛脱氢酶1（ALDH1）蛋白的表达以及细胞多向分化能力对该干细胞模型的可靠性进行鉴定。给予10-10 mol/L 17β-雌二醇(E2β)对该干细胞模型处理7 d，观察上述相关指标的变化。结果：去除胰岛素的悬浮培养方法获得的瘤细胞球呈葡萄状，由30～60个细胞组成。细胞球显示细胞角蛋白18和CD10蛋白均表达阳性，CD44+CD24-细胞的含量及ALDH1蛋白表达量均较贴壁细胞明显增高（P<0.05）。使用10-10 mol/L E2β处理MCF7肿瘤球细胞7 d，肿瘤球细胞数及体积增大。使用10-10 mol/L E2β处理MCF7肿瘤球细胞24 h，CD44+CD24-细胞数量及ALDH1蛋白表达量较未处理组明显升高（P<0.05）。结论：去除胰岛素的悬浮培养方法可有效建立乳腺癌干细胞体外研究模型，并可用于E2β对乳腺癌干细胞生长调控的研究。

Breast cancer stem cells cultured without insulin and effect of estrogen induction

AIM:To determine whether suspension culture medium without insulin can be used to feed breast cancer tumorsphere, or not. METHODS:MCF7 cells were used to build tumorsphere. The morphological changes, CD44+ CD24- expression, aldehyde dehydrogenase 1 (ALDH1) expression and multiple division ability were measured to identify the breast cancer stem cells and to detect the function of 17β-estradiol (E2β) in tumorsphere of MCF7 cells. RESULTS:The tumorshere, each containing 30 to 60 cells, was obtained by the method of insulin-removal suspension culture. These cells were cytokeratin 18 and CD10 proteins positive, and the number of CD44+ CD24- cells and ALDH1 protein expression were significantly higher than the adherent cultured cells (P<0.05). Using 10-10  mol/L E2β to treat the tumorshere for 7 d, the tumor cell number and volume were significantly increased. Using 10-10  mol/L E2β to treat the tumorshere for 24 h, the CD44+ CD24-cells and ALDH1 protein expression were significantly higher than those in non-treatment group (P<0.05). CONCLUSION: Suspension culture medium without insulin can be used to feed breast cancer tumorsphere. These tumorsphere could be used as a model to determine the function of E2β in breast cancer stem cell research.