花青素抑制肾细胞癌的肿瘤发生。

Cyanidin Curtails Renal Cell Carcinoma Tumorigenesis.

作者信息

Liu Xiaobing, Zhang Dangling, Hao Yaxing, Liu Qian, Wu Yuqi, Liu Xin, Luo Jing, Zhou Tao, Sun Bishao, Luo Xing, Xu Jie, Wang Qingqing, Yang Zhenxing, Li Longkun

机构信息

Department of Urology, Xinqiao Hospital, Third Military Medical University, Chongqing, China.

Institute of Immunology, Third Military Medical University, Chongqing, China.

出版信息

Cell Physiol Biochem. 2018;46(6):2517-2531. doi: 10.1159/000489658. Epub 2018 May 5.

DOI:10.1159/000489658

PMID:29742507

Abstract

BACKGROUND/AIMS: Cyanidin is an anthocyanin found in many foods. Although its variable antioxidant levels are well-documented, little is known about its effects on renal cell carcinoma (RCC) tumorigenesis. This study, therefore, investigated the effects of cyanidin on the proliferation, migration, and invasion of renal cell carcinoma lines and demonstrated, for the first time, significant inhibitory effects of cyanidin on RCC tumorigenesis.

METHODS

RCC cells were treated with different doses of cyanidin and the effects were tested by Cell Counting Kit-8 reagent, clone formation assay, transwell assay, and flow cytometry. Moreover, the cyanidin-mediated mechanism that curtailed tumorigenesis was analyzed by RNA sequencing (RNA-seq). Sequencing data from The Cancer Genome Atlas (TCGA) were used to compare the expression of both early growth response protein 1 (EGR1) and selenoprotein W (SEPW1) in RCC and tumor-free adjacent normal tissue samples. Real-time PCR (RT-PCR) and/or western blot were used to assess the expression of E-cadherin, cleaved-caspase3, Bcl2, p62, and ATG4.

RESULTS

We found significantly greater induction of cell-cycle arrest, apoptosis, and suppression of RCC cell invasion and migration at concentrations of 25 µM and 100 µM than at a concentration of 50 µM. It was also discovered, first through RNA-seq then confirmed by RT-PCR, that cyanidin (100 µM) inhibited RCC carcinogenesis through EGR1 and SEPW1. TCGA data indicated that the expression level of EGR1 was lower and that of SEPW1 was higher in RCC tumor tissue than in normal tissues. Moreover, western blot and/or RT-PCR indicated that cleaved-caspase3 was enhanced and E-cadherin was inhibited by cyanidin treatment. Furthermore, western blot and RT-PCR also showed regulation of p62 and ATG4, which are associated with autophagy. Cyanidin in vivo significantly inhibited the growth of xenografts in nude mice.

CONCLUSIONS

The results of this study showed the therapeutic potential of cyanidin for the treatment of RCC and the prevention of recurrence and metastasis.

摘要

背景/目的：花青素是一种存在于多种食物中的花色苷。尽管其抗氧化水平的变化已有充分记录，但关于其对肾细胞癌（RCC）肿瘤发生的影响知之甚少。因此，本研究调查了花青素对肾癌细胞系增殖、迁移和侵袭的影响，并首次证明了花青素对RCC肿瘤发生具有显著抑制作用。

方法

用不同剂量的花青素处理RCC细胞，并通过细胞计数试剂盒-8试剂、克隆形成试验、Transwell试验和流式细胞术检测其效果。此外，通过RNA测序（RNA-seq）分析花青素介导的抑制肿瘤发生的机制。利用来自癌症基因组图谱（TCGA）的测序数据比较RCC组织和癌旁正常组织样本中早期生长反应蛋白1（EGR1）和硒蛋白W（SEPW1）的表达。采用实时荧光定量PCR（RT-PCR）和/或蛋白质免疫印迹法评估E-钙黏蛋白、裂解的半胱天冬酶3、Bcl-2、p62和自噬相关蛋白4（ATG4）的表达。

结果

我们发现，与50 μM浓度相比，25 μM和100 μM浓度的花青素对细胞周期阻滞、细胞凋亡的诱导作用以及对RCC细胞侵袭和迁移的抑制作用显著更强。还发现，首先通过RNA-seq，然后经RT-PCR证实，花青素（100 μM）通过EGR1和SEPW1抑制RCC的发生。TCGA数据表明，RCC肿瘤组织中EGR1的表达水平低于正常组织，而SEPW1的表达水平高于正常组织。此外，蛋白质免疫印迹法和/或RT-PCR表明，花青素处理可增强裂解的半胱天冬酶3的表达，并抑制E-钙黏蛋白的表达。此外，蛋白质免疫印迹法和RT-PCR还显示了与自噬相关的p62和ATG4的表达受到调控。花青素在体内显著抑制裸鼠异种移植瘤的生长。