Lu Cheng-Hsin, Wu Chun-Hsien, Hsieh Pei-Fang, Wu Chen-Yu, Kuo Wade Wei-Ting, Ou Chien-Hui, Lin Victor Chia Hsiang
Division of Urology, Penghu Hospital, Penghu 880001, Taiwan, R.O.C.
Division of Urology, Department of Surgery, E-Da Hospital, Kaohsiung 824005, Taiwan, R.O.C.
Oncol Lett. 2022 Mar;23(3):90. doi: 10.3892/ol.2022.13210. Epub 2022 Jan 21.
Enzalutamide is one of the options for treating patients with castration-resistant or metastatic prostate cancer. However, a substantial proportion of patients become resistant to enzalutamide after a period of treatment. Cells in these tumors typically exhibit increased proliferative and migratory capabilities, in which N-cadherin (CDH2) appear to serve an important role. In the present study, by up- and downregulating the expression of CDH2, the possible effects of CDH2 on the prostate cancer cell line LNCaP were investigated. Male sex hormone-sensitive LNCaP cells treated with 10 µM enzalutamide were named LNCaP enzalutamide-resistant (EnzaR) cells. Reverse transcription-PCR, western blotting and immunofluorescence staining were used to measure CDH2, E-cadherin, α-SMA, Snail and Slug expression. Transfection with the pCMV-CDH2 plasmid was performed for CDH2 upregulation, whilst transfection with small interfering RNA (siRNA)-CDH2 was performed for CDH2 downregulation. MTT and Cell Counting Kit-4 assays were used to evaluate the proportion of viable cancer cells. Subsequently, gap closure assay was performed to evaluate the migratory capability of both LNCaP and LNCaP EnzaR cell lines. CDH2 expression was found to be increased in LNCaP EnzaR cells compared with that in LNCaP cells. CDH2 overexpression increased cell viability and migration in both LNCaP and LNCaP EnzaR cell lines. By contrast, the opposite trend was observed after CDH2 expression was knocked down. CDH2 expression also showed a high association with that of four epithelial-mesenchymal transition markers, which was confirmed by western blotting. Based on these results, it was concluded that knocking down CDH2 expression using siRNA transfection mediated significant influence on LNCaP EnzaR cell physiology, which may be a potential therapeutic option for prostate cancer treatment.
恩杂鲁胺是治疗去势抵抗性或转移性前列腺癌患者的选择之一。然而，相当一部分患者在经过一段时间的治疗后会对恩杂鲁胺产生耐药性。这些肿瘤中的细胞通常表现出增殖和迁移能力增强，其中N-钙黏蛋白（CDH2）似乎发挥着重要作用。在本研究中，通过上调和下调CDH2的表达，研究了CDH2对前列腺癌细胞系LNCaP的可能影响。用10 μM恩杂鲁胺处理的雄激素敏感LNCaP细胞被命名为LNCaP恩杂鲁胺耐药（EnzaR）细胞。采用逆转录聚合酶链反应、蛋白质印迹法和免疫荧光染色法检测CDH2、E-钙黏蛋白、α-平滑肌肌动蛋白、Snail和Slug的表达。通过转染pCMV-CDH2质粒上调CDH2表达，同时通过转染小干扰RNA（siRNA）-CDH2下调CDH2表达。采用MTT法和细胞计数试剂盒-4检测法评估存活癌细胞的比例。随后，进行划痕愈合试验以评估LNCaP和LNCaP EnzaR细胞系的迁移能力。发现LNCaP EnzaR细胞中CDH2的表达高于LNCaP细胞。CDH2过表达增加了LNCaP和LNCaP EnzaR细胞系的细胞活力和迁移能力。相比之下，在敲低CDH2表达后观察到相反的趋势。蛋白质印迹法证实，CDH2表达也与四种上皮-间质转化标志物的表达高度相关。基于这些结果，得出结论：通过siRNA转染敲低CDH2表达对LNCaP EnzaR细胞生理学有显著影响，这可能是前列腺癌治疗的一种潜在治疗选择。
