Zhang Zhenghao, Xu Wenfang, Ye Kewen, Lv Shanmei, Wu Jintao, Zhou Yadi
Clinical Laboratory, Affiliated Hospital of Shaoxing University, Shaoxing, China.
Clinical Laboratory, Shaoxing People's Hospital, Shaoxing, China.
Transl Androl Urol. 2025 Apr 30;14(4):897-912. doi: 10.21037/tau-2025-11. Epub 2025 Apr 27.
Piperlongumine (PL) is a natural alkaloid obtained from the long pepper and can inhibit the progression of various tumors. However, its role in renal cell carcinoma (RCC) remains unclear. Thus, the purpose of this study was to determine whether PL can suppress RCC progression and to clarify the related mechanisms.
Cell Counting Kit-8 (CCK-8) and colony formation assays were applied to characterize the effect of PL in RCC cell proliferation; meanwhile, cellular reactive oxygen species (ROS), Fe level, and mitochondrial membrane potential (MMP) assays were used to determine PL's role in mitochondrial homeostasis. Immunofluorescence experiments using MitoTracker red was applied to visualize the morphology of mitochondria. Western blotting and coimmunoprecipitation (Co-IP) and RNA immunoprecipitation (RNA-IP) assays were used to examine PL's regulation of DDX11 expression via miR-15b-3p. In addition, a xenograft mouse model was created to clarify the effect of DDX11 overexpression on RCC progression with or without PL treatment.
We found that PL inhibited RCC cell proliferation in a dose-dependent manner by interfering with the mitochondrial homeostasis of RCC cells. In terms of mechanism, RNA sequencing showed that PL decreased the expression of DDX11, inhibited the maturation of miR-15b-3p, and further increased the level of dihydrolipoamide dehydrogenase (DLD) to disrupt the mitochondrial homeostasis of RCC cells. In addition, the vivo xenograft mouse model showed that DDX11 plays a stimulatory role in PL-induced RCC inhibition.
Our study demonstrated that DDX11 contributes to PL-induced RCC inhibition by modulating the miR-15b-3p-DLD axis.
胡椒碱（PL）是一种从长胡椒中提取的天然生物碱，能够抑制多种肿瘤的进展。然而，其在肾细胞癌（RCC）中的作用仍不清楚。因此，本研究的目的是确定PL是否能抑制RCC进展并阐明相关机制。
采用细胞计数试剂盒-8（CCK-8）和集落形成试验来表征PL对RCC细胞增殖的影响；同时，使用细胞活性氧（ROS）、铁水平和线粒体膜电位（MMP）试验来确定PL在线粒体稳态中的作用。使用MitoTracker红进行免疫荧光实验以观察线粒体的形态。采用蛋白质免疫印迹法、免疫共沉淀（Co-IP）和RNA免疫沉淀（RNA-IP）试验来检测PL通过miR-15b-3p对DDX11表达的调控。此外，建立异种移植小鼠模型以阐明在有或无PL治疗的情况下DDX11过表达对RCC进展的影响。
我们发现PL通过干扰RCC细胞的线粒体稳态以剂量依赖的方式抑制RCC细胞增殖。在机制方面，RNA测序表明PL降低了DDX11的表达，抑制了miR-15b-3p的成熟，并进一步提高了二氢硫辛酰胺脱氢酶（DLD）的水平，从而破坏了RCC细胞的线粒体稳态。此外，体内异种移植小鼠模型表明DDX11在PL诱导的RCC抑制中起促进作用。
我们的研究表明，DDX11通过调节miR-15b-3p-DLD轴促进PL诱导的RCC抑制。
