Abstract
The prognosis of bladder urothelial carcinoma (BLCA) varies greatly even for patients with similar pathological characteristics. We conducted transcriptome sequencing on ten pairs of BLCA samples and adjacent normal tissues to identify differentially expressed genes. Anillin (ANLN) was identified as a transcript that was significantly up-regulated in BLCA samples compared with normal tissues. Prognostic power of candidate gene was studied using qRT-PCR and immunohistochemistry on 40 and 209 patients, respectively. Patients with elevated ANLN expression level was correlated with poorer cancer-specific (median, 22.4 vs. 37.3 months, p = 0.001), progression-free (median, 19.7 vs. 27.9 months, p = 0.001) and recurrence-free survival (median, 17.1 vs. 25.2 months, p = 0.011) compared with low ANLN expression. Public datasets TCGA and NCBI-GEO were analyzed for external validation. Knockdown of ANLN in J82 and 5637 cells using small interfering RNA significantly inhibited cell proliferation, migration, and invasion ability. Moreover, knockdown of ANLN resulted in G2/M phase arrest and decreased expression of cyclin B1 and D1. Microarray analysis suggested that ANLN played a major role in cell migration and was closely associated with several cancer-related signaling pathways. In conclusion, ANLN was identified as a promising prognostic biomarker which could be used to stratify different risks of BLCA.
Highlights
Bladder urothelial carcinoma (BLCA) is the sixth most common cancers in North America, and the sixth leading cause of cancer-related deaths in Europe[1, 2]
The cutoff of log2-fold change >1 and probability >0.6, a method proposed by Tarazona et al.[18] which was more effective in controlling the rate of false discoveries, was used to filter genes that were differentially expressed between bladder urothelial carcinoma (BLCA) samples and the matched normal controls
RNA-Seq analysis of ten pairs of BLCA samples identified 1516 genes that were differentially expressed between tumor and normal tissue
Summary
Bladder urothelial carcinoma (BLCA) is the sixth most common cancers in North America, and the sixth leading cause of cancer-related deaths in Europe[1, 2]. Identification of prognostic molecular markers is critical to help urologists in the stratification of highand low-risk BLCA patients to make individualized treatment decisions[7, 8]. Transcriptome sequencing (RNA-seq) of tumors is an efficient approach to identify molecules involved in carcinogenesis and reveal biomarkers with prognostic value[3, 12]. We performed RNA-Seq analysis on ten pairs of BLCA samples and their matched adjacent normal tissue. We observed a number of differentially expressed genes common to the ten pair samples. Among these genes, we found ANLN expression was correlated with pathological stage and was a promising prognostic biomarker. Our data suggested that ANLN was a novel and promising prognostic biomarker for BLCA that may aid in the risk stratification
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