Gene expression of SUM159-mir100 ALDH+ and ALDH- cells from CTRL or mir100-overexpressing group. Homo sapiens

Emerging evidence suggest that miRNAs play an essential role in self-renewal and differentiation of normal and malignant stem cells by regulating the expression of key stem cell regulatory genes. Here we demonstrate that mir-100 expression is related to cellular differentiation state with lowest expression in cells displaying stem cell markers. Utilizing a tetracycline inducible lentivirus driving mir-100 expression, we found that mir-100 overexpression decreased breast cancer stem cells (BCSCs) and inhibited cancer cell proliferation in vitro and in mouse xenografts by targeting SMARCA5, SMARCD1 and BMPR2. Overall design: In order to determine the cellular targets of mir-100 in BCSCs and non-BCSCs, ALDH-positive and -negative populations of pTRIPZ-SUM159-mir-100 cells were separated and cultured in suspension for ten hours in the presence or absence of DOX. Gene expression profiles of the four populations were determined utilizing Affymetrix microarrays

越来越多的证据表明，微小RNA（microRNAs，miRNAs）通过调控关键干细胞调控基因的表达，在正常干细胞与恶性干细胞的自我更新及分化过程中发挥核心作用。本研究证实，miR-100的表达水平与细胞分化状态密切相关，在表达干细胞标志物的细胞中其表达量最低。本研究使用四环素诱导型慢病毒驱动miR-100表达，发现miR-100过表达可通过靶向SMARCA5、SMARCD1及BMPR2，在体外及小鼠异种移植瘤模型中减少乳腺癌干细胞（breast cancer stem cells, BCSCs）的数量并抑制癌细胞增殖。整体实验设计：为明确miR-100在乳腺癌干细胞及非乳腺癌干细胞中的细胞靶点，本研究分离得到pTRIPZ-SUM159-miR-100细胞的乙醛脱氢酶阳性（aldehyde dehydrogenase, ALDH）与阴性群体，分别在存在或不存在多西环素（doxycycline, DOX）的条件下进行悬浮培养10小时。随后通过Affymetrix基因芯片检测这四个群体的基因表达谱。