Berberine Maintains the Neutrophil N1 Phenotype to Reverse Cancer Cell Resistance to Doxorubicin.

Shuhui Zhang,Mengqi Wang,Lin Zhou,Yuehua Wang,Gangjun Du,Mengdi Zhang,Jincheng Du,Shengnan Geng,Fuguang Kui,Jiahuan Li,Wenwen Gu

doi:10.3389/fphar.2019.01658

Abstract

This study explores the contributions of neutrophils to chemotherapeutic resistance and berberine-regulated cancer cell sensitivity to doxorubicin (DOX). In vitro experiments, continuous DOX treatment led to the shift of HL-60 cells to N2 neutrophils and thus induced chemotherapeutic resistance. The combination treatment with DOX and 2 µM berberine resulted in the differentiation of HL-60 cells toward N1 and therefore stimulated HL-60 cell immune clearance. Berberine increased reactive oxygen species (ROS) and decreased autophagy and therefore induced apoptosis in HL-60-N2 cells with morphological changes, but had no effect on cell viability in HL-60-N1 cells. The neutrophil-regulating efficacy of berberine was confirmed in the urethane-induced lung carcinogenic model and H22 liver cancer allograft model. Furthermore, we found that DOX-derived neutrophils had high levels of CD133 and CD309 surface expression, which prevented both chemotherapeutic sensitivity and immune rejection by self-expression of PD-L1 and surface expression of PD-1 receptor on T cells, whereas berberine could downregulate CD133 and CD309 surface expression. Finally, berberine-relevant targets and pathways were evaluated. This study first suggests an important role of berberine in regulating neutrophil phenotypes to maintain cancer cell sensitivity to DOX.

Highlights

Cancer is a leading cause of human disease-related death, and chemotherapy is currently the standard of care (Chi et al, 2019; Matei et al, 2019)
There was a significant difference between HL-60-N1 and HL-60-N2 cells (Figure 1F); simultaneously, HL-60-N2 cells had a reduction in reactive oxygen species (ROS), as indicated by intracellular fluorescence of DCFH-DA (Figure 1G), and an increase in autophagy, as indicated by more LC3B and the less p62 immunofluorescence (Figure 2A)
Systemic cancer chemotherapy exerts extensive cytotoxic effects and results in immunogenic or nonimmunogenic cell death of tumor and normal tissue (Verwaal et al, 2003; Kono et al, 2013; Kuijpers et al, 2014). These excessive dead cells are beyond the scavenging capacity of professional and non-professional neighboring phagocytes and are able to mimic pathogen response-like recruitment of neutrophils to maintain unresolved inflammation for cancer initiation and progression (Garg et al, 2017)

Summary

Introduction

Cancer is a leading cause of human disease-related death, and chemotherapy is currently the standard of care (Chi et al, 2019; Matei et al, 2019). Cancer cells usually respond to initial chemotherapy but become resistant and increase metastatic potential, which greatly contributes to mortality from most types of cancer (Berns and Berns, 2017; Wojtaszek et al, 2019). In this sense, cancer chemotherapy can stimulate tumor spread if cancer cells cannot be completely eliminated. Tumor cell persistence requires resistance to cell death and immune escape in cancer bearing hosts (Hanahan and Weinberg, 2019; Ponzetta et al, 2019); the relationship between chemotherapeutic resistance and immune responses has received little attention

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Conclusion