SIRT3 促进作用通过调节 FOXO3/CDT1 轴降低肺癌对顺铂的耐药性。

SIRT3 promotion reduces resistance to cisplatin in lung cancer by modulating the FOXO3/CDT1 axis.

机构信息

The Third Department of Medical Oncology, the Third People's Hospital of Zhengzhou, Zhengzhou, P. R. China.

Medical Department, Women & Infants Hospital of Zhengzhou, Zhengzhou, P. R. China.

出版信息

Cancer Med. 2021 Feb;10(4):1394-1404. doi: 10.1002/cam4.3728.

DOI:10.1002/cam4.3728

PMID:33655712

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7926010/

Abstract

BACKGROUND

Cisplatin is an extensively used chemotherapy agent for lung cancer, but its drug resistance serves as a huge obstacle for chemotherapy failure of lung cancer patients. Hence, researchers aimed to determine role of sirtuin 3 (SIRT3) considering its action in cisplatin resistance of lung cancer.

METHODS

The expression patterns of SIRT3, FOXO3, and CDT1 were determined using RT-qPCR and Immunoblotting in lung cancer. Immunofluorescence and Co-IP were adopted to detect co-localization and interaction of FOXO3 and CDT1. Loss- and gain-function assays were conducted to determine roles of SIRT3, FOXO3, and CDT1 in resulting pathological changes, while biological behavior of cells was determined using a combination of CCK-8, flow cytometry, colony formation, and Transwell assays. The effects of SIRT3 and CDT1 were determined in the nude mice xenografted with the tumor. The proliferation-, angiogenesis-, and apoptosis-associated factors levels were determined using Immunoblotting.

RESULTS

SIRT3, FOXO3, and CDT1 expression was suppressed in the lung cancer tissues and cells. FOXO3 positively regulates the CDT1 expression pattern and SIRT3 elevation inhibits FOXO3 at the acetylated level, thus, elevating FOXO3 expression. The elevation of SIRT3, FOXO3, or CDT1 inhibited cell cisplatin resistance of lung cancer cells as well as inhibited viability, proliferation, and invasion in vitro. In vivo experiments, SIRT3 depletion elevated Ki-67 and VEGFA levels, but downregulated cleaved caspase 3 level.

CONCLUSION

Collectively, overexpressed SIRT3 elevates expression of FOXO3a/CDT1 axis, thus, contributing to enhanced sensitivity of lung cancer cells.

摘要

背景

顺铂是一种广泛应用于肺癌的化疗药物，但它的耐药性是肺癌患者化疗失败的巨大障碍。因此，研究人员旨在确定 SIRT3（沉默信息调节因子 3）的作用，因为它可以影响肺癌的顺铂耐药性。

方法

通过 RT-qPCR 和免疫印迹法测定肺癌中 SIRT3、FOXO3 和 CDT1 的表达模式。采用免疫荧光和 Co-IP 检测 FOXO3 和 CDT1 的共定位和相互作用。通过失活和增益功能实验确定 SIRT3、FOXO3 和 CDT1 在导致病理变化中的作用，同时使用 CCK-8、流式细胞术、集落形成和 Transwell 实验组合确定细胞的生物学行为。通过裸鼠移植肿瘤来确定 SIRT3 和 CDT1 的作用。通过免疫印迹法测定增殖、血管生成和凋亡相关因子水平。

结果

SIRT3、FOXO3 和 CDT1 的表达在肺癌组织和细胞中受到抑制。FOXO3 正向调节 CDT1 的表达模式，而 SIRT3 的升高抑制 FOXO3 的乙酰化水平，从而升高 FOXO3 的表达。SIRT3、FOXO3 或 CDT1 的升高抑制了肺癌细胞对顺铂的耐药性，并抑制了体外细胞的活力、增殖和侵袭。体内实验中，SIRT3 耗竭升高了 Ki-67 和 VEGFA 的水平，但降低了 cleaved caspase 3 的水平。