Zhang Fangxi, Wan Xiaofeng, Zhan Jianmin, Shen Ming, Li Runsheng
National Health Commission (NHC) Key Lab of Reproduction Regulation (Shanghai Institute for Biomedical and Pharmaceutical Technologies), School of Pharmacy, Fudan University, Shanghai, China.
Department of Pharmacy and Examination, Heze Medical Collge, Heze, China.
Front Oncol. 2024 Mar 7;14:1361152. doi: 10.3389/fonc.2024.1361152. eCollection 2024.
Prostate cancer (PCa) is the second most common solid cancer among men worldwide and the fifth leading cause of cancer-related deaths in men. Sulforaphane (SFN), an isothiocyanate compound, has been shown to exert inhibitory effects on a variety of cancers. However, the biological function of SFN in PCa has not been fully elucidated. The objective of this study was conducted to further investigate the possible underlying mechanism of SFN in PCa using cell culture and in vivo tumor model experiments.
Cell viability, migration, invasion, and apoptosis were analyzed by Cell Counting Kit-8 (CCK-8), wound healing assay, transwell assay, or flow cytometry. Expression of microRNA (miR)-3919 was detected by quantitative real-time polymerase chain reaction (qRT-PCR) or in situ hybridization assay. Xenograft assay was conducted to validated the antitumor effect of miR-3919. The targeting relationship between miR-3919 and DJ-1 was verified by dual-luciferase reporter assay. The level of DJ-1was measured by qRT-PCR or western blotting (WB).
In the present study, SFN downregulated mRNA and protein expression of DJ-1, an oncogenic gene. Small RNA sequencing analysis and dual-luciferase reporter assay confirmed that microRNA (miR)-3919 directly targeted DJ-1 to inhibition its expression. Furthermore, miR-3919 overexpression impeded viability, migration, and invasion and promoted apoptosis of PCa cells. Tumor growth in nude mice was also inhibited by miR-3919 overexpression, and miR-3919 expression in PCa tissues was lower than that in peritumoral tissues in an hybridization assay. Transfection with miR-3919 inhibitors partially reversed the effects of SFN on cell viability, migration, invasion, and apoptosis.
Overall, the miR-3919/DJ-1 axis may be involved in the effects of SFN on the malignant biological behavior of PCa cells, which might be a new therapeutic target in PCa.
前列腺癌（PCa）是全球男性中第二常见的实体癌，也是男性癌症相关死亡的第五大主要原因。萝卜硫素（SFN）是一种异硫氰酸酯化合物，已被证明对多种癌症具有抑制作用。然而，SFN在PCa中的生物学功能尚未完全阐明。本研究的目的是通过细胞培养和体内肿瘤模型实验进一步探讨SFN在PCa中可能的潜在机制。
采用细胞计数试剂盒-8（CCK-8）、伤口愈合试验、Transwell试验或流式细胞术分析细胞活力、迁移、侵袭和凋亡。通过定量实时聚合酶链反应（qRT-PCR）或原位杂交试验检测微小RNA（miR）-3919的表达。进行异种移植试验以验证miR-3919的抗肿瘤作用。通过双荧光素酶报告基因试验验证miR-3919与DJ-1之间的靶向关系。通过qRT-PCR或蛋白质印迹法（WB）检测DJ-1的水平。
在本研究中，SFN下调了致癌基因DJ-1的mRNA和蛋白质表达。小RNA测序分析和双荧光素酶报告基因试验证实，微小RNA（miR）-3919直接靶向DJ-1以抑制其表达。此外，miR-3919过表达阻碍了PCa细胞的活力、迁移和侵袭，并促进了其凋亡。miR-3919过表达也抑制了裸鼠体内的肿瘤生长，并且在原位杂交试验中，PCa组织中miR-3919的表达低于瘤周组织。用miR-3919抑制剂转染部分逆转了SFN对细胞活力、迁移、侵袭和凋亡的影响。
总体而言，miR-3919/DJ-1轴可能参与了SFN对PCa细胞恶性生物学行为的影响，这可能是PCa的一个新的治疗靶点。
