Comprehensive analyses reveal the carcinogenic and immunological roles of ANLN in human cancers

Abstract

Background: Anillin (ANLN) is an actin-binding protein that is essential for cell division and contributes to cell growth and migration. Although previous studies have shown that ANLN is related to carcinogenesis, no pan-cancer analyses of ANLN have been reported. Accordingly, in this study, we evaluated the carcinogenic roles of ANLN in various cancer types using online databases.

Methods: We evaluated the potential carcinogenic roles of ANLN using TIMER2 and Gene Expression Omnibus databases with 33 types of cancers. We further investigated the associations of ANLN with patient prognosis, genetic alterations, phosphorylation levels, and immune infiltration in multiple cancers using GEPIA2, cBioPortal, UACLAN, and TIMER2 databases. Additionally, the potential functions of ANLN were explored using Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses. Reverse transcription quantitative polymerase chain reaction and immunohistochemistry were used to determine ANLN mRNA and protein expression in colorectal cancer (CRC), gastric cancer (GC), and hepatocellular carcinoma (HCC) cell lines.

Results: ANLN was overexpressed in various tumor tissues compared with corresponding normal tissues, and significant correlations between ANLN expression and patient prognosis, genetic alterations, phosphorylation levels, and immune infiltration were noted. Moreover, enrichment analysis suggested that ANLN functionally affected endocytosis, regulation of actin cytoskeleton, and oxytocin signaling pathways. Importantly, ANLN mRNA and protein expression levels were upregulated in gastrointestinal cancers, including CRC, GC, and HCC.

Conclusions: Our findings suggested that ANLN participated in tumorigenesis and cancer progression and may have applications as a promising biomarker of immune infiltration and prognosis in various cancers.

Keywords: Anillin; Biomarker; Cancer; Immune infiltration; Prognosis.

Fig. 1

ANLN expression and its correlation with clinicopathological features. A The expression of ANLN mRNA in human normal testis tissues. B The expression of ANLN mRNA in human normal bone marrow tissues. C Analysis ANLN expression in various tumors or certain tumor subtypes in TIMER2 database. D Analysis of ACC, DLBC, LAML, LGG, OV, SARC, TGCT, and UCS expression between TCGA tumor tissues and GTEx database the corresponding normal tissues. E Analysis of ANLN total protein expression between breast cancer, clear cell RCC, colon cancer, ovarian cancer, LUAD, UCEC and corresponding normal tissues in CPTAC dataset. F Analysis of ANLN in pathological stages of ACC, BLCA, CESC, COAD, KICH, LUAD, PAAD, THCA. *P < 0.05; **P < 0.01; ***P < 0.001; Log scale adopts Log2 (TPM + 1)

Fig. 2

The correlation analysis between ANLN expression and prognosis of cancers in TCGA database using GEPIA2. A Overall survival. B Disease-free survival. The survival map with positive results were presented

Fig. 3

The correlation between ANLN expression and prognosis of certain cancers using the Kaplan–Meier plotter containing OS, RFS, PFS, PPS, DMFS, FP, and DFS. A Breast cancer. B Gastric cancer. C Lung cancer. D Ovarian cancer. E Liver cancer

Fig. 4

Analysis of ANLN genetic alteration of TCGA cancers the cBioPortal database. A The alteration frequency with mutation type. B The alteration frequency with mutation site. C The correlation between alteration status and prognosis in UCEC and LUNG

Fig. 5

Analysis of ANLN protein phosphorylation in cancers using CPTAC of UACLAN database. A Phosphorylation sites with significant differences are marked in ANLN protein (NP_061155.2, S6, S102, S182, S225, S272, S485, S518, S562, S664, S755, S792, S800 sites). B Expression of different ANLN protein phosphorylation sites in breast cancer. C Expression of ANLN protein phosphorylation sites S182 and S485 in LUAD, ovarian cancer and UCEC. D Expression of ANLN phosphorylation sites S225, S755, S792 and S518 in cancers

Fig. 6

Analysis of the correlation between ANLN expression and immune cells. A CD4⁺T cells. B CD8⁺T cells. C Macrophage. D NK cells. E B cells. F Neutrophils

Fig. 7

Analysis of the correlation between ANLN expression and immune cell infiltration of cancer-associated fibroblasts in cancers. A The heatmap of all tumors in TCGA generated by EPIC, MCPCUNTER, and TIDE algorithm. B The scatter plots of cancers generated by one algorithm, including BLCA, BLCA-LumA, ESCA, HNSC, LUAD, HNSC-HPV-, TGCT, and MESO

Fig. 8

Enrichment analysis ANLN-related genes. A The interaction network of 50 ANLN-binding proteins with available experimentally evidenced in the STING. B The correlation between ANLN and selected genes, including GPR62, PLP1, MAG, TMEM144, RAGGCP1 using GEPIA2. C The corresponding heatmap in the all cancers are presented. D Venn diagram of the ANLN-binding and related genes. E Analysis of KEGG pathway. F The cnetplot for the molecular function based on GO analysis

Fig. 9

Validation of ANLN mRNA and protein expression in gastrointestinal malignancies. A–C qRT-PCR analysis detected the ANLN mRNA expression relative to GAPDH in colorectal cancer, gastric cancer, and hepatocellular carcinoma cell lines. D–G The Human Protein Atlas database validated the ANLN protein expression in liver cancer, colorectal cancer, stomach cancer, pancreatic cancer and corresponding normal tissues. *P < 0.05