CENPF was downregulated in LUAD and LUSC
To evaluate the CENPF expression in TCGA, we analyzed CENPF expression base on the RNA-seq data of kinds of malignancies cancers. The differential expression between the tumor and normal tissues for CENPF in all TCGA tumors is shown in Figure.1A. CENPF expression was significantly higher in BLCA (bladder urothelial carcinoma), CHOL (cholangiocarcinoma) ,COAD (colon adenocarcinoma), ESCA(Esophageal carcinoma),HNSC(head and neck cancer),KICH (kidney chromophobe), KIRC (kidney renal clear cell carcinoma),KIRP(Kidney renal papillary cell carcinoma), LIHC (liver hepatocellular carcinoma),LUAD (lung adenocarcinoma),LUSC(Lung squamous cell carcinoma) compared with adjacent or normal tissues. In addition, the express level of CENPF was significantly higher in LUAD (lung adenocarcinoma), LUSC (Lung squamous cell carcinoma) is shown in Figure.1B, C.
Prognostic role of CENPF in LUAD and LUSC
Further, In order to elucidate the Prognostic role of CENPF in LUAD and LUSC. When CENPF expression level was divided into top 50% versus lower 50%, we found that higher CENPF (top 50% vs bottom 50%) expression was significantly associated with worsened OS and DFS in LUAD (Figure 2A, B). We then explored whether CENPF expression was associated with survival in LUSC, we found that higher CENPF expression was significantly associated with improved OS and DFS (Figure 2C，D).
Prognostic role of immune Infiltration Level in LUAD and LUSC
To explore the prognostic role of immune infiltration level in LUAD and LUSC. The B cells, CD8+ T cells, CD4+ T cells, macrophage, neutrophils, and dendritic cells (DCs) was analyzed in the prognostic role. Accordingly, increased infiltrating levels of B cells, macrophage was significantly associated with improved prognosis in LUAD (Figure 3A). We then explored whether immune infiltration level was associated with survival in LUSC cohort and found that higher B cells, and CD8+ T cells, macrophages, neutrophils, and dendritic cells (DCs) infiltrating levels was significantly associated with improved survival (Figure 3B).
Association of CENPF with cancer immunity in LUAD and LUSC
We therefore hypothesized that immunity in LUAD and LUSC could be linked to CENPF expression. Cases with higher CENPF expression showed significantly increased infiltration of B cells, CD8+T cells in LUAD (Figure 4A) and increased infiltration of B cells, and CD8+ T cells, macrophages, neutrophils, and dendritic cells in LUSC (Figure 4B).We analyzed correlation between CENPF expression and tumor immune infiltration level abundance indicated by decreased tumor purity. We found that higher CENPF expression was significantly associated with increased purity only in LUSC (Figure 4C). We then illustrated whether CENPF copy number between tumor immune infiltration level abundance. We found that CENPF copy number was significantly associated with increased tumor immune infiltration level (B cells, and CD4+ T cells, macrophages, neutrophils, and dendritic cells) in LUAD (Figure 4D). In the LUSC, CENPF copy number was significantly associated with increased tumor immune infiltration level (B cells, CD8+ T and CD4+ T cells, macrophages, neutrophils, and dendritic cells) in LUAD (Figure 4E).
CENPF and cancer immunity marker in LUAD and LUSC
As higher PD-L1, PD1, CTLA4 was associated with worsened prognosis in lung cancers, we hypothesized that our findings were due to increased cancer immunity with concomitant increased PD-L1, PD1, CTLA4 expression in LUAC and LUSC. Specifically, PD-L1 expression was associated with increased CD8+ cells, neutrophils, and B cells in LUAD and LUSC (Figure 5A, B). In LUAD cohort, we found that CENPF expression was significantly correlated with increased PD-L1(r=0.113, P= 0.010) but not with PD-1 and CTLA4 expression (P=0.081, P=0.343). Also, CENPF expression was significantly correlated with increased PD1(r= 0.1148, P=0) and CTLA4(r=-0.15, P=0) but not with PD-L1 (P=0.755) in LUSC cohort.
CENPF knockdown suppresses the proliferation, migration and invasion of NSCLC cells
We next determined the effects of CENPF knockdown on the proliferation potential of NSCLC cells in vitro. Our CCK8 assay results showed that CENPF knockdown group inhibited the growth abilities of H1299 and A549 cells than that of control group (Figure 6A). In addition, the results of colony formation assays showed that CENPF knockdown also inhibited the clone formation abilities of H1299 and A549 cells (Figure 5B). In order to find whether CENPF is related to the cancer metastasis in NSCLC cells, H1299 and A549 cells were seeded in the culture palte for cell scratch assay to detect cell migration ability. The results indicated that CENPF knockdown group inhibited the migration of H1299 and A549 cells (Figure 6C). Further, the transwell migration and invasion assay was used to analyze the function of CENPF in NSCLC cells migration and invasion. It was also demonstrated that CENPF knockdown inhibited the migration and invasion of H1299 and A549 cell (Figure 6D).