萝卜硫素作为依维莫司的辅助药物可对抗肾癌细胞系中的依维莫司耐药性。

Sulforaphane as an adjunctive to everolimus counteracts everolimus resistance in renal cancer cell lines.

作者信息

Juengel Eva, Euler Stephanie, Maxeiner Sebastian, Rutz Jochen, Justin Saira, Roos Frederik, Khoder Wael, Nelson Karen, Bechstein Wolf O, Blaheta Roman A

机构信息

Department of Urology, Goethe-University, Interdisciplinary Science Building, Building 25A, Room 404, Theodor-Stern-Kai 7, Frankfurt am Main, D-60590, Germany.

Department of Vascular and Endovascular Surgery, Goethe-University, Frankfurt am Main, Germany.

出版信息

Phytomedicine. 2017 Apr 15;27:1-7. doi: 10.1016/j.phymed.2017.01.016. Epub 2017 Jan 31.

DOI:10.1016/j.phymed.2017.01.016

PMID:28314474

Abstract

BACKGROUND

The mechanistic target of rapamycin (mTOR) inhibitors, everolimus and temsirolimus, have widened therapeutic options to treat renal cell carcinoma (RCC). However, chronic treatment with these inhibitors often induces resistance, leading to therapeutic failure.

PURPOSE

The natural compound, sulforaphane (SFN), was added to an everolimus based regime in vitro in the hopes of preventing resistance development.

METHODS

A panel of RCC cell lines (A498, Caki-1, KTCTL-26) was treated with everolimus or SFN or with an everolimus-SFN-combination, either short- (24h) or long-term (8 weeks), and cell growth, proliferation, apoptosis, and cell cycle phases were measured. The cell cycle regulating proteins cdk1, cdk2, cyclin A, cyclin B, akt and raptor (both total and activated) were also evaluated.

RESULTS

Short-term incubation with everolimus (1nM) or SFN (5µM) significantly reduced RCC cell growth. Additive effects on tumor growth and proliferation were evoked by the SFN-everolimus combination. Long-term everolimus-incubation led to resistance development in Caki-1 cells, evidenced by elevated growth and proliferation, associated with an increased percentage of G2/M (non-synchronized cell model) or S-phase (synchronized cell model) cells. Molecular analysis revealed up-regulation of the cdk1-cyclin B and cdk2-cyclin A axis, along with elevated phosphorylation of the mTOR sub-member, raptor. In contrast, resistance development was not observed with the long-term combination of SFN-everolimus. The combination suppressed Caki-1 growth and proliferation, and was associated with an increase in G0/G1-phase cells, diminished cdk1 and akt (both total and activated), cyclin B and raptor expression.

CONCLUSION

Adding SFN to an everolimus based RCC treatment regimen in vitro delayed resistance development observed with chronic everolimus monotherapy. Ongoing in vivo studies are necessary to verify the in vitro data.

摘要

背景

雷帕霉素的作用靶点（mTOR）抑制剂依维莫司和替西罗莫司拓宽了肾细胞癌（RCC）的治疗选择。然而，长期使用这些抑制剂常常会诱导耐药性，导致治疗失败。

目的

将天然化合物萝卜硫素（SFN）添加到基于依维莫司的治疗方案中，以期在体外预防耐药性的产生。

方法

使用依维莫司或SFN或依维莫司与SFN的组合，对一组肾癌细胞系（A498、Caki-1、KTCTL-26）进行短期（24小时）或长期（8周）处理，并检测细胞生长、增殖、凋亡和细胞周期阶段。还评估了细胞周期调节蛋白cdk1、cdk2、细胞周期蛋白A、细胞周期蛋白B、akt和猛禽（总蛋白和活化蛋白）。

结果

短期用依维莫司（1nM）或SFN（5µM）孵育可显著降低肾癌细胞的生长。SFN与依维莫司的组合对肿瘤生长和增殖产生相加效应。长期用依维莫司孵育导致Caki-1细胞产生耐药性，表现为生长和增殖增加，G2/M期（非同步细胞模型）或S期（同步细胞模型）细胞百分比增加。分子分析显示cdk1-细胞周期蛋白B和cdk2-细胞周期蛋白A轴上调，同时mTOR亚基猛禽的磷酸化增加。相比之下，长期使用SFN与依维莫司的组合未观察到耐药性的产生。该组合抑制了Caki-1的生长和增殖，并与G0/G1期细胞增加、cdk1和akt（总蛋白和活化蛋白）、细胞周期蛋白B和猛禽表达减少有关。