背景:已发现apelin受体(APLNR)的异常表达与各种癌症的发展有关,然而,其在前列腺癌(PCa)中的功能尚不清楚。本研究旨在探讨APLNR在PCa中的作用及潜在机制。
方法:采用qRT-PCR检测APLNRmRNA的表达。通过平板克隆和流式细胞术测定PCa细胞的增殖和凋亡。此外,凋亡相关蛋白的表达(Bax,使用蛋白质印迹评估Bcl-2和裂解的caspase-3)。通过免疫荧光和蛋白质印迹分析DNA损伤标记(γ-H2AX)。进行GSEA分析以寻求PCa中APLNR的富集途径,和PI3K的蛋白质水平,p-PI3K,AKT,p-AKT,mTOR,和p-mTOR使用蛋白质印迹进行测试。
结果:APLNR表达在PCa组织和细胞中上调。沉默APLNR增强PCa细胞对放疗的敏感性,表现为抑制细胞增殖,促进细胞凋亡,促进DNA损伤。接下来,沉默APLNR抑制PI3K/AKT/mTOR通路。具体来说,740Y-P(PI3K/AKT/mTOR通路激活剂)逆转沉默APLNR对PCa细胞增殖的影响,细胞凋亡和DNA损伤。
结论:沉默APLNR抑制细胞增殖,促进细胞凋亡,增强了PCa细胞的放射敏感性,参与PI3K/AKT/mTOR信号通路。本研究有利于加深对PCa的认识,进一步为PCa的治疗提供新的视角。
BACKGROUND: Aberrant expression of apelin receptor (APLNR) has been found to be involved in various cancers\' development, however, its function in prostate cancer (PCa) remains unclear. The research aimed to investigate the role and potential mechanism of APLNR in PCa.
METHODS: The mRNA expression of APLNR was detected via qRT-PCR assay. PCa cell proliferation and apoptosis were determined through plate cloning and flow cytometry. In addition, the expression of apoptosis-related proteins (Bax, Bcl-2, and cleaved caspase-3) was evaluated using western blot. DNA damage marker (γ-H2AX) was analyzed by immunofluorescence and western blot. GSEA analysis was performed for seeking enrichment pathways of APLNR in PCa, and the protein levels of PI3K, p-PI3K, AKT, p-AKT, mTOR, and p-mTOR were tested using western blot.
RESULTS: APLNR expression was up-regulated in PCa tissues and cells. Silencing APLNR enhanced the sensitivity of PCa cells to radiotherapy, which was manifested by inhibiting cell proliferation, promoting cell apoptosis, and promoting DNA damage. Next, silencing APLNR inhibited the PI3K/AKT/mTOR pathway. Specifically, 740Y-P (the PI3K/AKT/mTOR pathway activator) reversed the effects of silencing APLNR on PCa cell proliferation, apoptosis and DNA damage.
CONCLUSIONS: Silencing APLNR inhibited cell proliferation, promoted cell apoptosis, and enhanced the radiosensitivity of PCa cells, which was involved in the PI3K/AKT/mTOR signaling pathway. This study is conducive to the deeper understanding of PCa and further provides a new perspective for the treatment of PCa.