Department of Medical Oncology, Harbin Medical University Cancer Hospital, Harbin, Heilongjiang, 150081, China.
Biotherapy Center, Harbin Medical University Cancer Hospital, Harbin, Heilongjiang, 150081, China.
Exp Mol Med. 2022 Aug;54(8):1290-1305. doi: 10.1038/s12276-022-00837-6. Epub 2022 Aug 30.
The functional role of microRNA-375 (miR-375) in the development of prostate cancer (PCa) remains controversial. Previously, we found that plasma exosomal miR-375 is significantly elevated in castration-resistant PCa (CRPC) patients compared with castration-sensitive PCa patients. Here, we aimed to determine how miR-375 modulates CRPC progression and thereafter to evaluate the therapeutic potential of human umbilical cord mesenchymal stem cell (hucMSC)-derived exosomes loaded with miR-375 antisense oligonucleotides (e-375i). We used miRNA in situ hybridization technique to evaluate miR-375 expression in PCa tissues, gain- and loss-of-function experiments to determine miR-375 function, and bioinformatic methods, dual-luciferase reporter assay, qPCR, IHC and western blotting to determine and validate the target as well as the effects of miR-375 at the molecular level. Then, e-375i complexes were assessed for their antagonizing effects against miR-375. We found that the expression of miR-375 was elevated in PCa tissues and cancer exosomes, correlating with the Gleason score. Forced expression of miR-375 enhanced the expression of EMT markers and AR but suppressed apoptosis markers, leading to enhanced proliferation, migration, invasion, and enzalutamide resistance and decreased apoptosis of PCa cells. These effects could be reversed by miR-375 silencing. Mechanistically, miR-375 directly interfered with the expression of phosphatase nonreceptor type 4 (PTPN4), which in turn stabilized phosphorylated STAT3. Application of e-375i could inhibit miR-375, upregulate PTPN4 and downregulate p-STAT3, eventually repressing the growth of PCa. Collectively, we identified a novel miR-375 target, PTPN4, that functions upstream of STAT3, and targeting miR-375 may be an alternative therapeutic for PCa, especially for CRPC with high AR levels.
miR-375(miR-375)在前列腺癌(PCa)发展中的功能作用仍存在争议。此前,我们发现与去势敏感型 PCa(CSPC)患者相比,去势抵抗型 PCa(CRPC)患者的血浆外泌体 miR-375 显著升高。在此,我们旨在确定 miR-375 如何调节 CRPC 的进展,然后评估负载 miR-375 反义寡核苷酸的人脐带间充质干细胞(hucMSC)衍生外泌体(e-375i)的治疗潜力。我们使用 miRNA 原位杂交技术评估 PCa 组织中 miR-375 的表达,通过增益和失能实验确定 miR-375 的功能,以及通过生物信息学方法、双荧光素酶报告基因检测、qPCR、免疫组化和 Western blot 确定和验证靶标以及 miR-375 在分子水平上的作用。然后,评估 e-375i 复合物对 miR-375 的拮抗作用。我们发现 miR-375 在 PCa 组织和癌症外泌体中的表达上调,与 Gleason 评分相关。强制表达 miR-375 增强了 EMT 标志物和 AR 的表达,但抑制了凋亡标志物的表达,导致 PCa 细胞增殖、迁移、侵袭和恩杂鲁胺耐药性增强,凋亡减少。这些作用可以通过 miR-375 沉默来逆转。从机制上讲,miR-375 直接干扰磷酸酶非受体型 4(PTPN4)的表达,进而稳定磷酸化 STAT3。应用 e-375i 可以抑制 miR-375,上调 PTPN4,下调 p-STAT3,最终抑制 PCa 的生长。总之,我们确定了一个新的 miR-375 靶标 PTPN4,它作为 STAT3 的上游发挥作用,靶向 miR-375 可能是 PCa 的一种替代治疗方法,特别是对于 AR 水平高的 CRPC。