Combinatorial treatment with IDX and cis inhibits tumor growth
along with increasing T cells infiltration.
(A) Representative mice and dissected tumors. For one mouse from the cis
group, where the inoculated tumor was deeply located inside the organ
tissue, that tumor could not be clearly dissected out and therefore was
not its representative image is not shown. (B) Tumor volume during
treatment. Tumor xenografts were established in athymic nude mice by
inoculation of C17 cancer cells. After the tumors had grown to
approximately 50 mm3, the animals were randomly divided into 6 groups (5
mice/group), which were treated every-other-day with 5 mg/kg cis, 10
mg/kg IDX, 20 mg/kg, 5 mg/kg cis plus 10 mg/kg IDX or 5 mg/kg cis plus
20 mg/kg IDX. Control mice received saline. Tumors were measured each
other day over a total of 21 days after cancer cell inoculation and
tumor volume was calculated. * p≤0.05 and ***p<0.0001 when
compared to control group. (C) Violin plot showing the percentage of
infiltrating cells expressing CD14+ (monocyte) and CD19+ (B cells) in
total live lymphocytes and cells expressing CD4+,
CD8+ and CD4+ CD8+in CD19- populations isolated from mice tumor,
measured by flow cytometry. Each point represents one mouse in each
group. ** p≤0.01 when compared to cis group. (D) Flow cytometric
analysis on tumor infiltrating CD8 + T cells upon aforementioned
treatment in total live lymphocytes. * p≤0.05 when compared to DMSO
group. (E) Expression of ENOX2 at single-cell resolution from a publicly
available sequencing cohort GSE150430. UMAP plot generated by using all
cells harvested from 15 patients. Epithelial clusters enriched with
mainly cells from tumor were circled and found to be the major cell type
expressing ENOX2. (F) UMAP plot of malignant cells from 11 patients.
Some patients including patient 11 and 13 were found to contain
malignant cells with high expression of ENOX2. (G) Representative tumor
histology staining of NPC clinical tissues for ENOX2 expression. (H)
Analysis of ENOX2 gene expression and methylation from four independent
public microarray datasets GSE34573, GSE53819, GSE12452 and GSE52068. *
p≤0.05 when compared to corresponding normal tissue.