Thiol oxidative stress-dependent degradation of transglutaminase2 via protein S-glutathionylation sensitizes 5-fluorouracil therapy in 5-fluorouracil-resistant colorectal cancer cells

Xia Li; Yan Ma; Junzhou Wu; Maowei Ni; Aiping Chen; Yun Zhou; Wumin Dai; Zhongjian Chen; Ruibin Jiang; Yutian Ling; Qinghua Yao; Wei Chen

doi:10.1016/j.drup.2023.100930

Thiol oxidative stress-dependent degradation of transglutaminase2 via protein S-glutathionylation sensitizes 5-fluorouracil therapy in 5-fluorouracil-resistant colorectal cancer cells

Drug Resist Updat. 2023 Mar:67:100930. doi: 10.1016/j.drup.2023.100930. Epub 2023 Jan 20.

Authors

Xia Li¹, Yan Ma², Junzhou Wu³, Maowei Ni³, Aiping Chen³, Yun Zhou², Wumin Dai³, Zhongjian Chen³, Ruibin Jiang⁴, Yutian Ling³, Qinghua Yao⁵, Wei Chen⁶

Affiliations

¹ The Cancer Hospital of the University of Chinese Academy of Sciences (Zhejiang Cancer Hospital), Hangzhou 310022, China; Institute of Basic Medicine and Cancer (IBMC), Chinese Academy of Sciences, Hangzhou 310022, China; Zhejiang Cancer Research Institute, The Cancer Hospital of the University of Chinese Academy of Sciences, Hangzhou 310022, China; Key Laboratory of Prevention, Diagnosis and Therapy of Upper Gastrointestinal Cancer of Zhejiang Province, Hangzhou 310022, China; Zhejiang Provincial Key Laboratory of Integrated Traditional Chinese and Western Medicine on Cancer, The Cancer Hospital of the University of Chinese Academy of Sciences, Hangzhou 310022, China.
² The Cancer Hospital of the University of Chinese Academy of Sciences (Zhejiang Cancer Hospital), Hangzhou 310022, China; Institute of Basic Medicine and Cancer (IBMC), Chinese Academy of Sciences, Hangzhou 310022, China.
³ The Cancer Hospital of the University of Chinese Academy of Sciences (Zhejiang Cancer Hospital), Hangzhou 310022, China; Institute of Basic Medicine and Cancer (IBMC), Chinese Academy of Sciences, Hangzhou 310022, China; Zhejiang Cancer Research Institute, The Cancer Hospital of the University of Chinese Academy of Sciences, Hangzhou 310022, China.
⁴ The Cancer Hospital of the University of Chinese Academy of Sciences (Zhejiang Cancer Hospital), Hangzhou 310022, China; Institute of Basic Medicine and Cancer (IBMC), Chinese Academy of Sciences, Hangzhou 310022, China; Zhejiang Cancer Research Institute, The Cancer Hospital of the University of Chinese Academy of Sciences, Hangzhou 310022, China; Key Laboratory of Prevention, Diagnosis and Therapy of Upper Gastrointestinal Cancer of Zhejiang Province, Hangzhou 310022, China.
⁵ The Cancer Hospital of the University of Chinese Academy of Sciences (Zhejiang Cancer Hospital), Hangzhou 310022, China; Institute of Basic Medicine and Cancer (IBMC), Chinese Academy of Sciences, Hangzhou 310022, China; Department of Integrated Chinese and Western Medicine, The Cancer Hospital of the University of Chinese Academy of Sciences, Hangzhou 310022, China; Zhejiang Provincial Key Laboratory of Integrated Traditional Chinese and Western Medicine on Cancer, The Cancer Hospital of the University of Chinese Academy of Sciences, Hangzhou 310022, China.
⁶ The Cancer Hospital of the University of Chinese Academy of Sciences (Zhejiang Cancer Hospital), Hangzhou 310022, China; Institute of Basic Medicine and Cancer (IBMC), Chinese Academy of Sciences, Hangzhou 310022, China; Zhejiang Cancer Research Institute, The Cancer Hospital of the University of Chinese Academy of Sciences, Hangzhou 310022, China; Key Laboratory of Prevention, Diagnosis and Therapy of Upper Gastrointestinal Cancer of Zhejiang Province, Hangzhou 310022, China; Zhejiang Provincial Key Laboratory of Integrated Traditional Chinese and Western Medicine on Cancer, The Cancer Hospital of the University of Chinese Academy of Sciences, Hangzhou 310022, China. Electronic address: chenwei@zjcc.org.cn.

PMID: 36736043
DOI: 10.1016/j.drup.2023.100930

Abstract

5-Fluorouracil (5-Fu) is a first-line drug for colorectal cancer (CRC) therapy. However, the development of 5-Fu resistance limits its chemotherapeutic effectiveness and often leads to poor prognoses of CRC. Transglutaminase 2 (TGM2), a member of the transglutaminase family, is considered to be associated with chemoresistance through apoptotic prevention in various cancers including CRC. TGM2 was found to be overexpressed in two 5-Fu-resistant CRC cell lines and down-regulated by increased thiol oxidative stress induced by inhibition of glutathione reductase (GR). The present study aimed to explore the role of TGM2 in 5-Fu-resistant CRC and the mechanism of action by which the elevated thiol oxidative stress down-regulates TGM2 protein level. The results revealed that 5-Fu-resistance induced by overexpression of TGM2 in CRC cells was reversed through up-regulation of thiol oxidative stress. Knockdown of TGM2 increased the chemosensitivity of CRC cells to 5-Fu. Thiol oxidative stress potentially enhanced the therapeutic effect of 5-Fu in the resistant CRC cells by promotion of 5-Fu-induced apoptosis through down-regulation of TGM2. The elevated thiol oxidative stress increased the S-glutathionylation of TGM2 and led to proteasomal degradation of TGM2. Furthermore, Cys193 was identified as the S-glutathionylation site in TGM2, and its mutation resulted in thiol oxidative stress-mediated CRC cell apoptotic resistance. TGM2-induced EMT was also suppressed by the elevated thiol oxidative stress. A xenograft tumor model confirmed the effect of thiol oxidative stress in the reversal of 5-Fu resistance in CRC cells in vivo. TGM2 protein expression level was found to be significantly higher in human CRC specimens than in non-cancerous colorectal tissues. Taken together, the present data suggest an important role of TGM2 in 5-Fu resistance in CRC cells. Up-regulation of thiol oxidative stress could be a potential therapeutic approach for treating 5-Fu-resistant CRC and TGM2 may serve as a potential therapeutic target of thiol oxidative stress.

Keywords: 5-Fluorouracil; Apoptosis; Colorectal cancer; S-glutathionylation; TGM2; Thiol oxidative stress.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cell Line, Tumor
Cell Proliferation
Colorectal Neoplasms* / drug therapy
Colorectal Neoplasms* / genetics
Colorectal Neoplasms* / pathology
Disease Models, Animal
Drug Resistance, Neoplasm / genetics
Fluorouracil / pharmacology
Fluorouracil / therapeutic use
Gene Expression Regulation, Neoplastic
Humans
MicroRNAs* / genetics
Oxidative Stress

Substances

Fluorouracil
MicroRNAs
TGM2 protein, human