Abstract
The immune checkpoint molecule B7 family member H4 (B7-H4) plays a similar role to programmed death-ligand 1 in tumor immune evasion by regulating T-cell-mediated immune responses. However, besides the role in T-cell immunity, B7-H4 also affects tumor cell biology by promoting tumor cell proliferation, metastasis and angiogenesis. In order to explore the effect of B7-H4 on tumor cell biology, it is necessary to investigate the gene expression profile when B7-H4 is overexpressed. In the present study, 786-O cells were transfected to stably express B7-H4. A microarray technique was subsequently used to screen B7-H4-related differentially expressed genes (DEGs) in B7-H4/786-O cells compared with negative control (NC)/786-O cells. The protein expression of the upregulated DEGs, including non-metastatic cells 5, NME/NM23 family member 5 (NME5), membrane metalloendopeptidase (MME), vascular non-inflammatory molecule 1 (VNN1), matrix metalloproteinase (MMP) 7, tumor necrosis factor, C-X-C motif chemokine ligand (CXCL) 8, CXCL1 and C-C motif chemokine ligand (CCL) 2, was investigated using western blotting. Kidney renal papillary cell carcinoma mRNA-sequencing data obtained from The Cancer Genome Atlas revealed that chemokines, including CXCL1/2/3, CXCL8, MMP7 and CCL20, were positively correlated with B7-H4 gene expression. Furthermore, 59 clinical renal cell carcinoma tissues were collected and analyzed by immunohistochemical staining. The results revealed the positive correlation of B7-H4 with CCL20 and CXCL8, and validated the DEGs identified in tumor cell lines. 786-O transfectants were inoculated into non-obese diabetic/severe combined immunodeficiency mice, and tumor growth was investigated. B7-H4 overexpression promoted tumor growth and administration of anti-CXCL8 antibody reversed this effect. Furthermore, B7-H4 overexpression increased the number of tumor-infiltrating neutrophils while inhibition of CXCL8 abrogated this effect. These data indicated that recruitment of neutrophils in the tumor microenvironment by CXCL8 serves an important role in the tumor promotion effect of B7-H4. The present study revealed a novel mechanism of B7-H4 in tumor promotion in addition to T cell inhibition.