Abstract：
BACKGROUND: Metastatic renal cell carcinoma (RCC) poses a huge challenge once it has become resistant to targeted therapy. Vasculogenic mimicry (VM) is a novel blood supply system formed by tumor cells that can circumvent molecular targeted therapies. As one of the herbal remedies, curcumin has been demonstrated to play antineoplastic effects in many different types of human cancers; however, its function and mechanism of targeting VM in RCC remains unknown.
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.
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.
摘要：
背景：转移性肾细胞癌（RCC）一旦对靶向治疗产生抗药性，就构成了巨大的挑战。血管生成拟态（VM）是由肿瘤细胞形成的新型血液供应系统，可以规避分子靶向治疗。作为草药之一，姜黄素已被证明在许多不同类型的人类癌症中发挥抗肿瘤作用；然而，其在RCC中靶向VM的功能和机制尚不清楚。
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。
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。
