https://orcid.org/0000-0002-6022-4714
Abstract
Introduction and Objective
The mitogen-activated protein kinase (MAPK) pathway is inhibited by the pan-target inhibitor Anlotinib, which induces tumor cell death. In addition to the common apoptosis and necrosis, there is also a pyroptosis mode of cancer cell death in recent years, which is mainly manifested by the cleavage of gasdermin proteins (GSDMs). Gasdermin B (GSDMB) participates in the progression and outcome of bladder cancer. The efficacy and mechanism of Anlotinib in the treatment of GSDMB-positive bladder tumors have not been studied to date.
Methods
The relationship between GSDMB expression and tumor stage, overall survival rate, immunotherapy response, tumor recurrence and progression rate was analyzed from the TCGA bladder cancer database. Anlotinib was used to treat GSDMB-positive bladder cancer in mice followed by flow analysis of the secretion of inflammatory factors related to pyroptosis and the level of anti-tumor factors. Western blot analysis detected which MAPK and MEK signal transduction pathways.
Results
TCGA data analysis showed that the overall survival rate of bladder cancer patients with high GSDMB expression was better than that of patients with low GSDMB expression. In vivo experiments showed that Anlotinib was more effective in the treatment of GSDMB-positive bladder cancer than GSDMB-negative bladder cancer. Anlotinib can increase the secretion of antitumor-related factors in GSDMB-positive bladder cancer such as TNF-a and CD107a. In addition, Anlotinib also induced an increase in GSDMB protein expression. Anlotinib treatment of GSDMB-positive bladder cancer decreased AKT and MEK protein expression, which were involved in Anlotinib signal transduction pathway.
Conclusion
Anlotinib has a strong antitumor effect on GSDMB-positive bladder tumors. This effect is mainly achieved by anlotinib stimulating the secretion of relevant antitumor factors by lymphocytes. The PI3K/AKT and MEK signal transduction pathways were inhibited by Anlotinib in bladder cancer expressing GSDMB protein.
Cell Lines
The 293T cell was purchased from National Collection of Authenticated Cell Cultures (Cat No. SCSP-502). The MB49 cell line was donated by Professor Lee Longcheng’s laboratory at University of California, San Francisco.Citation20 The use of the MB49 cell was approved by the Ethics Committee of Xinhua Hospital.
Statement of Ethics
All the research was conducted ethically in accordance with the World Medical Association Declaration of Helsinki. The animal study protocol was approved by the Ethics Committee of Xinhua Hospital. Animal experiments were performed in accordance with the Health Guidelines of the National Institutes for the Care and Use of Laboratory Animals. The human data from TCGA (publicly available database) has been approved by the Ethics Committee of Xinhua Hospital Affiliate to Shanghai Jiaotong University School of Medicine.
Acknowledgments
We appreciate Chia Tai Tianqing Company for providing Anlotinib.
Disclosure
The authors have no conflicts of interest to declare for this work.