RNA-sequencing Reveal the Molecular Mechanism of Moxibustion on Neutrophil Gene Expression Profile During Breast Cancer Chemotherapy

Background: Moxibustion is a Traditional Chinese Medicine (TCM) therapy that can prevent neutropenia after breast cancer (BC) chemotherapy. We aimed to explore the mechanism of moxibustion for treating BC chemotherapy-induced neutropenia (CIN).MethodsFifty patients with BC undergoing chemotherapy and who met the inclusion criteria were randomized into an intervention (IT) group and control (OC) group. After excluding cases, there were 14 cases in the RNA sequencing (RNA-seq) group and 17 cases in the verification group. Neutrophils were extracted from the peripheral blood of the patients in both groups before and after chemotherapy. RNA-seq and bioinformatics analysis were performed in RNA-seq group. The verification group were verified using real time quantity polymerase chain reaction (RT-qPCR).ResultsRNA-seq screened 1092 DEGs before and after chemotherapy in the OC group, and 571 DEGs in the IT group. Compared with the OC group, the IT group had 707 DEGs after chemotherapy. The effect of moxibustion on the patients’ neutrophil gene expression profiles was related to the cell adhesion, adaptive immune response and metabolic pathways, and leukocyte migration, etc. The co-network results showed that TSC22D1 and TGFB1I1 have core regulatory function. GO and KEGG enrichment pathway analysis suggested a close correlation with the TGFβ1/TSC22D1-mediated cell adhesion molecules (CAMs) pathway. RT-qPCR showed that CD177 in the neutrophils was significantly upregulated after chemotherapy compared with that before chemotherapy (P < 0.05) without moxibustion, while TSC22D1 showed a downward trend (P = 0.094). Moxibustion significantly increased the expression of TSC22D1, ANKFY1, and ITGB3 in the neutrophils (P < 0.05) after BC chemotherapy.ConclusionThe mechanism of moxibustion in improving CIN may be related to the regulation of the TSC22D1 expression profile in neutrophil. We present a novel insight into the mechanism of moxibustion treatment of CIN in patients with BC.Trial registrationChinese Clinical Trial Register, ChiCTR-INR-16009557. Registered 23 October 2016, http://www.chictr.org.cn/showproj.aspx?proj=16203.