Immunology Research Center, Tabriz University of Medical Sciences, Daneshghah Ave, Tabriz, Iran.
Department of Cancer and Inflammation Research, Institute of Molecular Medicine, University of Southern Denmark, Odense, Denmark.
Mol Biol Rep. 2020 May;47(5):3691-3703. doi: 10.1007/s11033-020-05411-9. Epub 2020 Apr 3.
One of the major barriers in cancer therapy is the resistance to conventional therapies and cancer stem cells (CSCs) are among the main causes of this problem. CD133 as a CSC marker displays stem cell-like properties, tumorigenic capacity, and drug resistance in various cancers. However, the molecular mechanism behind CD133 function in prostate cancer (PC) still remains unclear. This research aimed to illustrate the probabilistic mechanism of CD133-siRNA and paclitaxel in the reduction of chemoresistance in PC cells. To measure the cell viability, migratory capacity, CSCs properties, invasive potential, apoptosis and cell cycle progression of the cells, the MTT, wound healing, spheroid assay, colony formation assay, DAPI staining and flow cytometry assays were applied in the LNCaP cell line, respectively. Also, quantitative real-time PCR (qRT-PCR) and western blot method were used for measuring the expression of CD133 and the effects of CD133 silencing on the AKT/mTOR/c-myc axis and pro-metastatic genes expression. We showed that the CD133-siRNA considerably decreased the CD133 expression. Moreover, CD133-siRNA and paclitaxel treatment significantly decreased cell proliferation and also inhibited the ability of cell migration and invasion and reduced pro-metastatic genes expression. Additionally, we found that the simultaneous use of CD133-siRNA and paclitaxel increased the paclitaxel-induced apoptosis. Our results confirmed that CD133 silencing combined with paclitaxel synergistically could suppress cell migration, invasion, and proliferation and enhance the chemosensitivity compared with mono treatment. Therefore, CD133 silencing therapy could be viewed as a promising and efficient strategy in PC targeted therapies.
癌症治疗的主要障碍之一是对常规疗法的耐药性,而癌症干细胞(CSC)是造成这一问题的主要原因之一。CD133 作为 CSC 标志物,在多种癌症中表现出干细胞样特性、致瘤能力和耐药性。然而,CD133 在前列腺癌(PC)中的功能的分子机制仍不清楚。本研究旨在阐明 CD133-siRNA 和紫杉醇降低 PC 细胞化疗耐药性的概率机制。为了测量细胞活力、迁移能力、CSC 特性、侵袭潜力、细胞凋亡和细胞周期进程,MTT、划痕愈合、球体形成试验、集落形成试验、DAPI 染色和流式细胞术分别应用于 LNCaP 细胞系。此外,还使用定量实时 PCR(qRT-PCR)和 Western blot 方法测量 CD133 的表达,以及 CD133 沉默对 AKT/mTOR/c-myc 轴和促转移基因表达的影响。结果显示,CD133-siRNA 可显著降低 CD133 的表达。此外,CD133-siRNA 和紫杉醇处理显著降低了细胞增殖,还抑制了细胞迁移和侵袭的能力,并降低了促转移基因的表达。此外,我们发现同时使用 CD133-siRNA 和紫杉醇可增加紫杉醇诱导的细胞凋亡。我们的结果证实,与单独治疗相比,CD133 沉默联合紫杉醇治疗可协同抑制细胞迁移、侵袭和增殖,增强化疗敏感性。因此,CD133 沉默疗法可作为 PC 靶向治疗的一种有前途和有效的策略。