ABSTRACT

LncRNA AFAP1-AS1 has been corroborated to function in diverse cancers. Our aim was to investigate the molecular mechanism of AFAP1-AS1 in PTX resistance in PCa. The levels of AFAP1-AS1, miR-195-5p, and FKBP1A were checked by qRT-PCR. 3-(4, 5-Dimethylthiazol-2-yl)-2, 5-Diphenyltetrazolium Bromide (MTT) assay was employed to assess the resistance of PTX-resistant PCa cells to PTX. Flow cytometry was introduced to evaluate cell apoptosis. The protein levels of C-caspase 3 were determined by western blot. The starBase was used to predict the interaction between miR-195-5p and AFAP1-AS1. Xenograft tumor model was established to investigate the biological role of AFAP1-AS1 in PTX resistance in vivo. The levels of AFAP1-AS1 and FKBP1A were upregulated in PCa tissues and cells, as well as PTX-resistant PCa cells, while the expression of miR-195-5p was declined. Knockdown of AFAP1-AS1 promoted the sensitivity of PTX-resistant PCa cells to PTX, induced apoptosis of PTX-resistant PCa cells, whereas the impacts could be reversed by reducing the expression of miR-195-5p. FKBP1A overexpression could rescue the effects of miR-195-5p-mediated enhancement on the sensitivity of PTX-resistant PCa cells to PTX, promotion on apoptosis of PTX-resistant PCa cells. AFAP1-AS1 interacted with miR-195-5p and miR-195-5p could bind to the 3ʹUTR of FKBP1A. AFAP1-AS1 silencing inhibited the tumor growth in mice implanted with PC3-TXR cell. The protein level of PCNA was decreased in PC3-TXR cells transfected with sh-AFAP1-AS1, while the expression of C-caspase 3 was upregulated. AFAP1-AS1 silencing attenuated the resistance of PTX-resistant PCa cells to PTX by downregulating FKBP1A via sponging miR-195-5p.

摘要

LncRNA AFAP1-AS1 已被证实在多种癌症中起作用。我们的目的是研究 AFAP1-AS1 在 PCa 中 PTX 抗性中的分子机制。通过 qRT-PCR 检查 AFAP1-AS1、miR-195-5p 和 FKBP1A 的水平。3-(4, 5-Dimethylthiazol-2-yl)-2, 5-Diphenyltetrazolium Bromide (MTT) 测定用于评估 PTX 抗性 PCa 细胞对 PTX 的抗性。引入流式细胞术来评估细胞凋亡。C-caspase 3 的蛋白质水平通过蛋白质印迹确定。starBase 用于预测 miR-195-5p 和 AFAP1-AS1 之间的相互作用。建立异种移植肿瘤模型研究AFAP1-AS1在体内PTX耐药中的生物学作用. AFAP1-AS1 和 FKBP1A 在 PCa 组织和细胞以及 PTX 抗性 PCa 细胞中的水平上调，而 miR-195-5p 的表达下降。AFAP1-AS1 的敲低促进了 PTX 抗性 PCa 细胞对 PTX 的敏感性，诱导了 PTX 抗性 PCa 细胞的凋亡，而这种影响可以通过降低 miR-195-5p 的表达来逆转。FKBP1A 过表达可以挽救 miR-195-5p 介导的增强对 PTX 抗性 PCa 细胞对 PTX 敏感性的影响，促进 PTX 抗性 PCa 细胞的凋亡。AFAP1-AS1 与 miR-195-5p 相互作用，miR-195-5p 可以与 FKBP1A 的 3ʹUTR 结合。AFAP1-AS1 沉默抑制了植入 PC3-TXR 细胞的小鼠的肿瘤生长。转染sh-AFAP1-AS1的PC3-TXR细胞中PCNA的蛋白水平降低，而 C-caspase 3 的表达上调。AFAP1-AS1 沉默通过海绵 miR-195-5p 下调 FKBP1A 来减弱 PTX 抗性 PCa 细胞对 PTX 的抗性。