YAP 的表达通过脂质过氧化诱导的铁死亡抑制视网膜母细胞瘤细胞增殖并提高化疗敏感性。

Department of Premiere Medical Service Center, The State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou, China.

Department of Oncology, Cancer Center, Integrated Hospital of Traditional Chinese Medicine, Southern Medical University, Guangzhou, China.

Chin Clin Oncol. 2023 Oct;12(5):52. doi: 10.21037/cco-23-97.

BACKGROUND

Retinoblastoma (RB) is a retinal cancer most commonly occurred in young children. Cisplatin and etoposide had been confirmed as chemotherapy drugs in the treatment of RB, even though the phenomenon of chemotherapeutic resistance has been occurring in clinical treatment frequently. RB has been reported to be a tumor with reduced expression of yes-associated protein (YAP). However, the role of YAP protein and its correlation with the chemotherapy effect in RB still remains unknown.

METHODS

Here we used human RB cell lines Y79 and RB3823 to construct YAP over-expression cell lines for exploring the specific role of YAP. In vitro, a series of techniques and methods were used to identify the biological role of YAP in RB, such as Agilent Seahorse assay, lipid peroxidation assay, intracellular reactive oxygen species (ROS) measurement, flow cytometry apoptosis assay, and other basic experimental techniques, among others.

RESULTS

The cell growth and cytology experimental results found YAP can inhibit the proliferation of RB cells and promote their apoptosis (Y79 32.71% vs. 3.75%; RB3823 40.32% vs. 6.73%). The mitochondrial fuel flex test, lipid peroxide and ROS measurement confirmed that YAP over-expression could promote mitochondrial fatty-acids β-oxidation and lipid peroxidation in RB cells. Quantitative real-time polymerase chain reaction (qRT-PCR) analysis for the expression of lipid peroxidation related factors imply that YAP over-expression caused ferroptosis in RB cell lines. In addition, YAP transcription specific activator PY-60 (10 µM) further improved the sensitivity of cisplatin/etoposide.

CONCLUSIONS

Our research results found the expression of YAP inhibits cell proliferation and promoted lipid peroxidation induced ferroptosis in RB. Interestingly, the mitochondrial oxidative phosphorylation shows an increased fatty acid dependency and decreased glucose dependency. As a result, this phenomenon improved the sensitivity of RB to cisplatin/etoposide chemotherapy in vitro. Our finding provides a potential therapeutic target for RB chemotherapy.

背景

视网膜母细胞瘤（RB）是一种常见于儿童的视网膜癌症。顺铂和依托泊苷已被确认为治疗 RB 的化疗药物，尽管在临床治疗中经常出现化疗耐药现象。已有报道称 RB 是一种 YAP 表达降低的肿瘤。然而，YAP 蛋白在 RB 中的作用及其与化疗效果的相关性尚不清楚。

方法

我们使用人 RB 细胞系 Y79 和 RB3823 构建 YAP 过表达细胞系，以探索 YAP 的特定作用。在体外，我们使用一系列技术和方法，如安捷伦 Seahorse 测定法、脂质过氧化测定法、细胞内活性氧（ROS）测量、流式细胞术凋亡测定法和其他基本实验技术，来鉴定 YAP 在 RB 中的生物学作用。

结果

细胞生长和细胞学实验结果发现 YAP 可以抑制 RB 细胞的增殖并促进其凋亡（Y79 为 32.71%比 3.75%；RB3823 为 40.32%比 6.73%）。线粒体燃料灵活性测试、脂质过氧化和 ROS 测量证实，YAP 过表达可促进 RB 细胞中线粒体脂肪酸β氧化和脂质过氧化。脂质过氧化相关因子的定量实时聚合酶链反应（qRT-PCR）分析表明，YAP 过表达导致 RB 细胞系发生铁死亡。此外，YAP 转录特异性激活剂 PY-60（10 μM）进一步提高了顺铂/依托泊苷的敏感性。