Sunnybrook Research Institute, University of Toronto, Toronto, Ontario, Canada.
Mol Cancer Res. 2013 May;11(5):443-55. doi: 10.1158/1541-7786.MCR-12-0461. Epub 2013 Feb 28.
Versican is highly expressed during the early stages of tissue development and its expression is elevated during wound repair and tumor growth. There is little literature on the potential role of breast cancer stem cells on the cellular-extracellular matrix interactions involving versican. An anti-versican short hairpin RNA (shRNA) was used to observe the effect of reduction of versican on breast cancer self-renewal. A versican G3 construct was exogenously expressed in breast cancer cell lines. Colony formation and mammosphere formation assays were conducted; flow cytometry was applied to analyze the prevalence of side population cells. The versican G3- and vector-transfected 66c14 cells were injected transdermally into BALB/c mice as a 10-fold dilution series from 1 × 10(5) to 1 × 10(2) cells per mouse. Versican G3 domain enhanced breast cancer self-renewal in both experimental in vitro and in vivo models. Versican G3-transfected cells contained high levels of side population cells, formed more mammospheres when cultured in the serum-free medium, and formed a greater number and larger colonies. Reduction of versican's functionality through anti-versican shRNA or knocking out the EGF-like motifs reduced the effect of versican on enhancing mammosphere and colony formation. Versican-enhanced self-renewal played a role in enhanced chemotherapeutic drug resistance, relating partly to the upregulated expression of EGF receptor (EGFR) signaling. Versican is highly expressed in breast cancer progenitor cells and was maintained at high levels before cell differentiation. Overexpression of versican enhanced breast cancer self-renewal through EGFR/AKT/GSK-3β (S9P) signaling and conferred resistant to chemotherapeutic drugs tested.
聚糖在组织发育的早期阶段高度表达，其表达在伤口修复和肿瘤生长过程中升高。关于乳腺癌干细胞对涉及聚糖的细胞-细胞外基质相互作用的潜在作用的文献很少。使用抗聚糖短发夹 RNA(shRNA)观察降低聚糖对乳腺癌自我更新的影响。外源性表达聚糖 G3 构建体在乳腺癌细胞系中。进行集落形成和乳腺球体形成测定;流式细胞术用于分析侧群细胞的流行率。将聚糖 G3 和载体转染的 66c14 细胞作为 10 倍稀释系列从每只小鼠 1×10(5)至 1×10(2)个细胞经皮注射到 BALB/c 小鼠中。聚糖 G3 结构域在实验体外和体内模型中均增强了乳腺癌的自我更新。转染聚糖 G3 的细胞含有高水平的侧群细胞，在无血清培养基中培养时形成更多的乳腺球体，并且形成更多和更大的集落。通过抗聚糖 shRNA 或敲除 EGF 样基序降低聚糖对增强乳腺球体和集落形成的作用来降低聚糖的功能。聚糖增强的自我更新在增强的化学治疗药物耐药性中起作用，部分与 EGF 受体(EGFR)信号的上调表达有关。聚糖在乳腺癌祖细胞中高度表达，并在细胞分化之前维持在高水平。聚糖的过表达通过 EGFR/AKT/GSK-3β(S9P)信号增强乳腺癌自我更新，并赋予对所测试的化学治疗药物的耐药性。
