GDC-0326 Enhances the Effects of 5-Fu in Colorectal Cancer Cells by Inducing Necroptotic Death

Zizhen Zhang; Fangyu Ju; Fei Chen; Haoyue Wu; Jingyu Chen; Jing Zhong; Liming Shao; Sheng Zheng; Liangjing Wang; Meng Xue

doi:10.2147/OTT.S302334

GDC-0326 Enhances the Effects of 5-Fu in Colorectal Cancer Cells by Inducing Necroptotic Death

Onco Targets Ther. 2021 Apr 13:14:2519-2530. doi: 10.2147/OTT.S302334. eCollection 2021.

Authors

Zizhen Zhang^#^{1

2

3}, Fangyu Ju^#^{1

2

3}, Fei Chen^{1

2

3}, Haoyue Wu⁴, Jingyu Chen^{1

2

3}, Jing Zhong^{1

2

3}, Liming Shao^{1

2

3}, Sheng Zheng^{1

2

3}, Liangjing Wang^{1

2

3}, Meng Xue^{1

2

3}

Affiliations

¹ Department of Gastroenterology, The Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou, 310020, People's Republic of China.
² Institution of Gastroenterology, Zhejiang University, Hangzhou, 310000, People's Republic of China.
³ Zhejiang University Cancer Center, Hangzhou, 310000, People's Republic of China.
⁴ Institute of Genetics and Department of Genetics, Zhejiang University School of Medicine, Hangzhou, 310012, People's Republic of China.

^# Contributed equally.

Abstract

Aim: Chemoresistance to 5-fluorouracil (5-Fu) is common in colorectal cancer (CRC). Programmed necrosis (necroptosis) is an alternative form of programmed cell death regulated by receptor-interacting protein kinase (RIPK) 1 and 3, assumed as a novel target of cancer therapy. In this study, we aimed to explore whether a novel small molecular agent GDC-0326 could facilitate the effect of 5-Fu through necroptosis.

Main methods: Cell Counting Kit-8 (CCK-8) assay and colony formation were performed to confirm the function of GDC-0326 in CRC cells. Western blot and immunofluorescence were conducted to measure the altered expressions of RIPK1/RIPK3 induced by GDC-0326. Subcutaneous tumor models were used to evaluate the chemotherapeutic effects and concomitant side effects of GDC-0326 in vivo.

Key findings: We found that GDC-0326 effectively suppressed the growth of CRC cells in a dose-dependent manner. The induction of necroptosis by GDC-0326 was correlated with the modulation of RIPK1 and RIPK3. Necrostatin-1 and GSK-872, inhibitors of RIPK1 and RIPK3, respectively, could rescue the cell death induced by GDC-0326. In addition, in vitro and in vivo studies showed that 5-Fu plus GDC-0326 evinced a better antitumor efficacy by suppressing tumor growth and increasing tumor necrosis with no increased toxicity.

Significance: This study demonstrates that GDC-0326 plus 5-Fu has augmented antitumor efficacy and acceptable safety, which might be a promising therapeutic strategy for CRC patients in the future.

Keywords: 5-fluorouracil; GDC-0326; colorectal cancer; combination therapy.

Grants and funding

This work was supported by the National Natural Science Foundation of China (grant nos. 82073229 and 81702308), Zhejiang Provincial Natural Science Foundation (grant nos. LY18H160009 and LY16H160031), and the National Natural Science Foundation of China (grant no. 81802348).