Knockdown of ZEB1 reverses cancer stem cell properties in prostate cancer cells
Knockdown of ZEB1 reverses cancer stem cell properties in prostate cancer cells
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DOI：
10.3892/or.2021.8009
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发表时间：
2021-05-01
期刊：
影响因子：
4.2
通讯作者：
Contreras, Hector R.
中科院分区：
文献类型：
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作者：
Perez, Gisella;Lopez-Moncada, Fernanda;Contreras, Hector R.
前列腺癌 (PCa) 是全球男性中第二常见的癌症类型。晚期前列腺癌对传统疗法有抵抗力，高复发率与高转移率相关。癌症干细胞（CSC）被认为是造成这一现象的原因，因为它们具有自我更新和分化成其他细胞类型的能力。锌指 E 盒结合同源盒 1 (ZEB1) 是一种参与上皮间质转化 (EMT) 调节的转录因子，与导致治疗耐药的多种机制的激活有关。最近的证据表明，CSC 可能在 EMT 期间起源于非 CSC，因此假设敲低 PCa 细胞系中的 ZEB1 表达可以恢复与 CSC 相关的一些特性。使用慢病毒转导，ZEB1 表达在 PCa DU145 和 LNCaP 细胞系中被沉默。分别使用逆转录定量 PCR 和蛋白质印迹分析测定关键经典 CSC 标志物（Kruppel 样因子 4、SOX2、CD44 和 CD133）的 mRNA 和蛋白表达水平。此外，还评估了ZEB1敲低细胞的集落形成能力，并对形成的集落类型（全克隆、旁克隆和分克隆）进行了表征。最后，在体外评估形成前列腺球的能力。结果发现，在ZEB1敲低的DU145细胞中，CSC表型标志物（CD44、CD133和SOX2）的表达水平较对照组降低。此外，ZEB1 敲低细胞形成前列腺球和产生集落的能力较低。总之，ZEB1 的稳定沉默逆转了 PCa 细胞系中的 CSC 特性。由于ZEB1与PCa细胞系中的恶性肿瘤、治疗耐药和CSC表型相关，因此靶向ZEB1可能是根除CSC并改善晚期PCa患者预后的关键因素。
Prostate cancer (PCa) is the second most diagnosed type of cancer in men worldwide. Advanced PCa is resistant to conventional therapies and high recurrence has been associated with high rates of metastasis. Cancer stem cells (CSCs) have been proposed to be responsible for this, due to their ability of self-renewal and differentiation into other cell types. Zinc finger E-box-binding homeobox 1 (ZEB1), a transcription factor involved in the regulation of epithelial-mesenchymal transition (EMT), has been associated with the activation of several mechanisms that lead to resistance to treatment. As recent evidence has shown that CSCs may originate from non-CSCs during EMT, it was hypothesized that knocking down ZEB1 expression in PCa cell lines could revert some properties associated with CSCs. Using lentiviraltransduction, ZEB1 expression was silenced in the PCa DU145 and LNCaP cell lines. The mRNA and protein expression levels of key canonical CSC markers (Kruppel-like factor 4, SOX2, CD44 and CD133) were determined using reverse transcription-quantitative PCR and western blot analysis, respectively. In addition, the colony forming ability of the ZEB1-knockdown cells was evaluated, and the type of colonies formed (holoclones, paraclones and meroclones) was also characterized. Finally, the ability to form prostatospheres was evaluated in vitro. It was found that in ZEB1-knockdown DU145 cells, the expression levels of CSC phenotype markers (CD44, CD133 and SOX2) were decreased compared with those in the control group. Furthermore, ZEB1-knockdown cells exhibited a lower ability to form prostatospheres and to generate colonies. In conclusion, stable silencing of ZEB1 reversed CSC properties in PCa cell lines. Since ZEB1 is associated with malignancy, therapy resistance and a CSC phenotype in PCa cell lines, targeting ZEB1 may be a key factor to eradicate CSCs and improve the prognosis of patients with advanced PCa.