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The LIM protein Ajuba promotes a metastatic phenotype in colon cancer cells

ABSTRACT: RNA-seq of Colon cancer cell line Sw480 with Ajuba OE and KD.Colon cancer, along with its potential for recurrence and metastasis is a major health problem. Understanding the proteins and pathways that regulate cell growth and metastasis may provide new targets for patient care. Ajuba is a LIM domain protein often found overexpressed in cancers, however the exact function role of Ajuba in colon cancer is not known. Our data demonstrates that Ajuba is highly expressed in colon cancer cell lines and tumour tissue compared to adjacent non-tumour tissue. Using publicly available data from The Cancer Genome Atlas, we correlated poor colon cancer patient survival with high Ajuba expression. To investigate the function of Ajuba in colon cancer metastasis we transduced cells with lentiviral constructs to knock down and overexpress Ajuba protein and performed an RNA-seq transcriptomic analysis on each modified line. Analysing the pathways enriched in the differentially expressed genes, we found Ajuba to be involved in cell proliferation, migration and differentiation. We confirmed our findings in biological assays, cells depleted of Ajuba were less proliferative, more sensitive to irradiation, migrated less and were less efficient in colony formation. Our data indicates that increased Ajuba expression observed in colon cancer supports proliferation, epithelial-to-mesenchymal transition and metastasis.

ORGANISM(S): Homo sapiens

PROVIDER: GSE147111 | GEO | 2020/07/25

REPOSITORIES: GEO

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Project description:Colorectal cancer (CRC) is one of the most prevalent cancers, with over one million new cases per year. Overall, prognosis of CRC largely depends on the disease stage and metastatic status. As precision oncology for patients with CRC continues to improve, this study aimed to integrate genomic, transcriptomic, and proteomic analyses to identify significant differences in expression during CRC progression using a unique set of paired patient samples while considering tumour heterogeneity.We analysed fresh-frozen tissue samples prepared under strict cryogenic conditions of matched healthy colon mucosa, colorectal carcinoma, and liver metastasis from the same patients. Somatic mutations of known cancer-related genes were analysed using Illumina's TruSeq Amplicon Cancer Panel; the transcriptome was assessed comprehensively using Clariom D microarrays. The global proteome was evaluated by liquid chromatography-coupled mass spectrometry (LC‒MS/MS) and validated by two-dimensional difference in-gel electrophoresis. Subsequent unsupervised principal component clustering, statistical comparisons, and gene set enrichment analyses were calculated based on differential expression results.Although panomics revealed low RNA and protein expression of CA1, CLCA1, MATN2, AHCYL2, and FCGBP in malignant tissues compared to healthy colon mucosa, no differentially expressed RNA or protein targets were detected between tumour and metastatic tissues. Subsequent intra-patient comparisons revealed highly specific expression differences (e.g., SRSF3, OLFM4, and CEACAM5) associated with patient-specific transcriptomes and proteomes.Our research results highlight the importance of inter- and intra-tumour heterogeneity as well as individual, patient-paired evaluations for clinical studies. In addition to changes among groups reflecting CRC progression, we identified significant expression differences between normal colon mucosa, primary tumour, and liver metastasis samples from individuals, which might accelerate implementation of precision oncology in the future.

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The LIM protein Ajuba promotes a metastatic phenotype in colon cancer cells

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