Wang Lina, Lin Yani, Meng Hui, Liu Chunyan, Xue Jing, Zhang Qi, Li Chaoyang, Zhang Pengju, Cui Fuai, Chen Weiwen, Jiang Anli
Department of Biochemistry and Molecular Biology, Shandong University School of MedicineJinan, P. R. China; Central Laboratory, The Second Hospital of Shandong UniversityJinan, P. R. China.
Department of Biochemistry and Molecular Biology, Shandong University School of Medicine Jinan, P. R. China.
Am J Transl Res. 2016 Dec 15;8(12):5219-5234. eCollection 2016.
The present study is to investigate the role of long non-coding RNAs (lncRNAs) in the development of androgen independence in prostate cancer and its underlying mechanism.
We established an androgen-independent prostate carcinoma (AIPC) cell line LNCaP-AI from androgen-dependent prostate carcinoma (ADPC) cell line LNCaP. Different expression profiles of lncRNAs and mRNAs between LNCaP and LNCaP-AI cells were investigated using microarray analysis. The expression of RNAs was determined using quantitative real-time polymerase chain reaction. Protein levels were measured using Western blotting. MTT assay was used to test cell viability. Tumor formation assay was performed in nude mice to detect tumor growth in vivo. Flow cytometry was performed to detect cell cycles. Transwell assay was employed to test cell migration and invasion.
According to bioinformatics prediction, lncRNA LOC283070 could possibly play an important role in the transition of LNCaP cells into LNCaP-AI cells. LOC283070 was up-regulated in LNCaP-AI cells and frequently up-regulated in AIPC cell lines. Overexpression of LOC283070 in LNCaP cells accelerated cell proliferation and migration, even under androgen-independent circumstances. Knockdown of LOC283070 inhibited LNCaP-AI cell proliferation and migration. Moreover, overexpression of LOC283070 promoted tumor growth in vivo in both normal mice and castrated mice. CAMK1D overexpression had similar effect with LOC283070, and CAMK1D knockdown fully abrogated the effect of LOC283070 overexpression on the transition of LNCaP cells into androgen-independent cells.
The present study shows that overexpression of LOC283070 mediates the transition of LNCaP cells into androgen-independent LNCaP-AI cells possibly via CAMK1D.
本研究旨在探讨长链非编码RNA（lncRNA）在前列腺癌雄激素非依赖性发展中的作用及其潜在机制。
我们从雄激素依赖性前列腺癌（ADPC）细胞系LNCaP建立了雄激素非依赖性前列腺癌细胞系LNCaP-AI。使用微阵列分析研究LNCaP和LNCaP-AI细胞之间lncRNA和mRNA的不同表达谱。使用定量实时聚合酶链反应测定RNA的表达。使用蛋白质印迹法测量蛋白质水平。使用MTT法测试细胞活力。在裸鼠中进行肿瘤形成试验以检测体内肿瘤生长。进行流式细胞术以检测细胞周期。采用Transwell试验测试细胞迁移和侵袭能力。
根据生物信息学预测，lncRNA LOC283070可能在LNCaP细胞向LNCaP-AI细胞的转变中起重要作用。LOC283070在LNCaP-AI细胞中上调，并且在AIPC细胞系中经常上调。在LNCaP细胞中过表达LOC283070可加速细胞增殖和迁移，即使在雄激素非依赖性情况下也是如此。敲低LOC283070可抑制LNCaP-AI细胞的增殖和迁移。此外，LOC283070的过表达促进了正常小鼠和去势小鼠体内的肿瘤生长。CAMK1D的过表达与LOC283070具有相似的作用，并且敲低CAMK1D完全消除了LOC283070过表达对LNCaP细胞向雄激素非依赖性细胞转变的影响。
本研究表明，LOC283070的过表达可能通过CAMK1D介导LNCaP细胞向雄激素非依赖性LNCaP-AI细胞的转变。
