OBJECTIVE: Here, in the work, we explored the role of curcumin and its molecular mechanism in the regulation of VM formation in RCC.
METHODS: RNA-sequencing analysis, immunoblotting, and immunohistochemistry were used to detect E Twenty Six-1(ETS-1), vascular endothelial Cadherin (VE-Cadherin), and matrix metallopeptidase 9 (MMP9) expressions in RCC cells and tissues. RNA sequencing was used to screen the differential expressed genes. Plasmid transfections were used to transiently knock down or overexpress ETS-1. VM formation was determined by tube formation assay and animal experiments. CD31-PAS double staining was used to label the VM channels in patients and xenograft samples.
RESULTS: Our results demonstrated that VM was positively correlated with RCC grades and stages using clinical patient samples. Curcumin inhibited VM formation in dose and time-dependent manner in vitro. Using RNA-sequencing analysis, we discovered ETS-1 as a potential transcriptional factor regulating VM formation. Knocking down or overexpression of ETS-1 decreased or increased the VM formation, respectively and regulated the expression of VE-Cadherin and MMP9. Curcumin could inhibit VM formation by suppressing ETS-1, VE-Cadherin, and MMP9 expression both in vitro and in vivo.
CONCLUSIONS: Our finding might indicate that curcumin could inhibit VM by regulating ETS-1, VE-Cadherin, and MMP9 expression in RCC cell lines. Curcumin could be considered as a potential anti-cancer compound by inhibiting VM in RCC progression.
目标:这里,在工作中,我们探讨了姜黄素在RCCVM形成调控中的作用及其分子机制。
方法:RNA测序分析,免疫印迹,和免疫组织化学用于检测E二十六-1(ETS-1),血管内皮钙粘蛋白(VE-Cadherin),基质金属肽酶9(MMP9)在RCC细胞和组织中的表达。RNA测序用于筛选差异表达基因。质粒转染用于瞬时敲低或过表达ETS-1。通过管形成测定和动物实验确定VM形成。CD31-PAS双重染色用于标记患者和异种移植物样品中的VM通道。
结果:我们的结果表明,使用临床患者样本,VM与RCC分级和分期呈正相关。姜黄素在体外以剂量和时间依赖性方式抑制VM形成。使用RNA测序分析,我们发现ETS-1是调节VM形成的潜在转录因子。敲除或过表达ETS-1减少或增加VM形成,分别调节VE-Cadherin和MMP9的表达。姜黄素可以通过抑制ETS-1、VE-Cadherin、和MMP9在体外和体内的表达。
结论:我们的发现可能表明姜黄素可以通过调节ETS-1,VE-Cadherin,和MMP9在RCC细胞系中的表达。姜黄素通过抑制VM在RCC进展中被认为是一种潜在的抗癌化合物。