文章：

SIRT1通过Beclin1去乙酰化介导的自噬促进膀胱癌顺铂耐药性

SIRT1 Promotes Cisplatin Resistance in Bladder Cancer via Beclin1 Deacetylation-Mediated Autophagy

原文发布日期：26 December 2023

DOI: 10.3390/cancers16010125

类型: Article

开放获取: 是

英文摘要：

Autophagy-dependent cisplatin resistance poses a challenge in bladder cancer treatment. SIRT1, a protein deacetylase, is involved in autophagy regulation. However, the precise mechanism through which SIRT1 mediates cisplatin resistance in bladder cancer via autophagy remains unclear. In this study, we developed a cisplatin-resistant T24/DDP cell line to investigate this mechanism. The apoptosis rate and cell viability were assessed using flow cytometry and the CCK8 method. The expression levels of the relevant RNA and protein were determined using RT-qPCR and a Western blot analysis, respectively. Immunoprecipitation was utilized to validate the interaction between SIRT1 and Beclin1, as well as to determine the acetylation level of Beclin1. The findings indicated the successful construction of the T24/DDP cell line, which exhibited autophagy-dependent cisplatin resistance. Inhibiting autophagy significantly reduced the drug resistance index of these cells. The T24/DDP cell line showed a high SIRT1 expression level. The overexpression of SIRT1 activated autophagy, thereby further promoting cisplatin resistance in the T24/DDP cell line. Conversely, inhibiting autophagy counteracted the cisplatin-resistance-promoting effects of SIRT1. Silencing SIRT1 led to increased acetylation of Beclin1, the inhibition of autophagy, and a reduction in the cisplatin resistance of the T24/DDP cell line. Introducing a double mutation (lysine 430 and 437 to arginine, 2KR) in Beclin-1 inhibited acetylation and activated autophagy, effectively reversing the decreased cisplatin resistance resulting from SIRT1 silencing. In summary, our study elucidated that SIRT1 promotes cisplatin resistance in human bladder cancer T24 cells through Beclin1-deacetylation-mediated autophagy activation. These findings suggest a potential new strategy for reversing cisplatin resistance in bladder cancer.

摘要翻译： 

自噬依赖性顺铂耐药是膀胱癌治疗中的一大挑战。SIRT1作为一种蛋白质去乙酰化酶，参与自噬调控过程。然而，SIRT1如何通过自噬介导膀胱癌顺铂耐药的具体机制尚不明确。本研究通过构建顺铂耐药T24/DDP细胞系，深入探讨了这一机制。我们采用流式细胞术和CCK8法分别检测细胞凋亡率和细胞活力，通过RT-qPCR和Western blot技术测定相关RNA及蛋白表达水平，并运用免疫共沉淀验证SIRT1与Beclin1的相互作用及Beclin1乙酰化水平。研究结果显示：成功构建的T24/DDP细胞系表现出自噬依赖性顺铂耐药特征，抑制自噬可显著降低该细胞系的耐药指数。T24/DDP细胞中SIRT1呈现高表达状态，过表达SIRT1能激活自噬并进一步促进T24/DDP细胞的顺铂耐药性；而抑制自噬则可抵消SIRT1的促耐药作用。沉默SIRT1会导致Beclin1乙酰化水平升高、自噬抑制及T24/DDP细胞顺铂耐药性降低。在Beclin-1中引入双突变（赖氨酸430和437突变为精氨酸，2KR）可抑制乙酰化并激活自噬，有效逆转因SIRT1沉默导致的顺铂耐药性下降。综上所述，本研究阐明SIRT1通过Beclin1去乙酰化介导的自噬激活途径，促进人膀胱癌T24细胞的顺铂耐药性，这为逆转膀胱癌顺铂耐药提供了潜在的新策略。

原文链接：

SIRT1 Promotes Cisplatin Resistance in Bladder Cancer via Beclin1 Deacetylation-Mediated Autophagy