Tang Xiaoshuang, Li Feng, Jia Jing, Yang Chao, Liu Wei, Jin Ben, Wang Xinyang, Gao Ruixia, He Dalin, Guo Peng
Department of Urology, The First Affiliated Hospital of Xi'an Jiaotong University.
Department of Urology, The Second Affiliated Hospital of Xi'an Jiaotong University.
Int J Nanomedicine. 2017 Apr 12;12:2979-2993. doi: 10.2147/IJN.S133009. eCollection 2017.
Androgen plays an important role in the progression of prostate cancer. In the present study, novel magnetic molecularly imprinted polymers (MMIPs) with good biocompatibility were produced for the selective separation and inhibition of testosterone in prostate cancer cells.
MMIPs were prepared by using magnetic nanospheres, gelatin, and testosterone as the supporting materials, functional monomer, and the template molecule, respectively. The characterization of the resultant products was investigated by transmission electron microscopy, X-ray diffraction, and vibrating sample magnetometry. To test whether MMIPs can remove testosterone in biologic samples, human LNCaP (androgen-dependent) and C4-2 (androgen-independent) prostate cancer cells were selected as cell models. The translocation of androgen receptor (AR) was detected by immunofluorescence assay, and the expression of PSA mRNA was detected by real-time quantitative polymerase chain reaction analysis. Cell flow cytometry analysis was performed to detect cell cycle arrest.
The synthesized nanomaterials (MMIPs) possessed high crystallinity, satisfactory superparamagnetic properties, and uniform imprinted shell, and exhibited high adsorption capacity, fast kinetics, and high selectivity for testosterone. Moreover, the obtained imprinted nanomaterials could selectively enrich and detect testosterone in the LNCaP cell samples as a solid-phase extractant coupled with high-performance liquid chromatography. In addition, the MMIPs could freely enter prostate cancer cells and suppress the translocation of AR into the cell nucleus. We further found that MMIPs inhibited upregulation of AR downstream target genes in LNCaP and C4-2 cells; also, MMIPs inhibited cell growth and induced obvious cell cycle arrest in androgen-dependent LNCaP cells, but had no obvious effect on androgen-independent C4-2 cells.
Our results indicate that the obtained imprinted nanomaterials can specifically and effectively bind testosterone and recover it from prostate cancer cells. Moreover, the MMIPs can freely enter prostate cancer cells and block the activation of testosterone-AR pathway. Thus, the MMIPs may be a new option for antiandrogen therapy in prostate cancer.
雄激素在前列腺癌进展中起重要作用。在本研究中，制备了具有良好生物相容性的新型磁性分子印迹聚合物（MMIPs），用于前列腺癌细胞中睾酮的选择性分离和抑制。
分别以磁性纳米球、明胶和睾酮为支撑材料、功能单体和模板分子制备MMIPs。通过透射电子显微镜、X射线衍射和振动样品磁强计对所得产物进行表征。为测试MMIPs是否能去除生物样品中的睾酮，选择人LNCaP（雄激素依赖型）和C4-2（雄激素非依赖型）前列腺癌细胞作为细胞模型。通过免疫荧光测定法检测雄激素受体（AR）的转位，通过实时定量聚合酶链反应分析检测PSA mRNA的表达。进行细胞流式细胞术分析以检测细胞周期阻滞。
合成的纳米材料（MMIPs）具有高结晶度、令人满意的超顺磁性特性和均匀的印迹壳，对睾酮表现出高吸附容量、快速动力学和高选择性。此外，所获得的印迹纳米材料作为与高效液相色谱联用的固相萃取剂，可选择性地富集和检测LNCaP细胞样品中的睾酮。此外，MMIPs可自由进入前列腺癌细胞并抑制AR向细胞核的转位。我们进一步发现，MMIPs抑制LNCaP和C4-2细胞中AR下游靶基因的上调；此外，MMIPs抑制雄激素依赖型LNCaP细胞的生长并诱导明显的细胞周期阻滞，但对雄激素非依赖型C4-2细胞无明显影响。
我们的结果表明，所获得的印迹纳米材料可以特异性且有效地结合睾酮并从前列腺癌细胞中回收它。此外，MMIPs可自由进入前列腺癌细胞并阻断睾酮-AR途径的激活。因此，MMIPs可能是前列腺癌抗雄激素治疗的新选择。
